Therapeutic compounds and methods of use thereof

ABSTRACT

The invention provides a compound of formula (I): 
                         
or a pharmaceutically acceptable salt thereof; wherein, A, B, ring C, R 3 , R 4 , R 5 , R 6 , and R 7  have the meaning as described herein, and compositions containing such compounds and methods for using such compounds and compositions.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority to International Application NumberPCT/CN2018/081250 filed Mar. 30, 2018. The entire content of theapplication referenced above is hereby incorporated by reference herein.

The present invention relates to organic compounds useful for therapy ina mammal, and in particular to inhibitors of sodium channel (e.g.,NaV1.7) that are useful for treating sodium channel-mediated diseases orconditions, such as pain, as well as other diseases and conditionsassociated with the modulation of sodium channels.

Voltage-gated sodium channels are transmembrane proteins that initiateaction potentials in nerve, muscle and other electrically excitablecells, and are a necessary component of normal sensation, emotions,thoughts and movements (Catterall, W. A., Nature (2001), Vol. 409, pp.988-990). These channels consist of a highly processed alpha subunitthat is associated with auxiliary beta subunits. The pore-forming alphasubunit is sufficient for channel function, but the kinetics and voltagedependence of channel gating are in part modified by the beta subunits(Goldin et al., Neuron (2000), Vol. 28, pp. 365-368).Electrophysiological recording, biochemical purification, and molecularcloning have identified ten different sodium channel alpha subunits andfour beta subunits (Yu, F. H., et al., Sci. STKE (2004), 253; and Yu, F.H., et al., Neurosci. (2003), 20:7577-85).

The sodium channel family of proteins has been extensively studied andshown to be involved in a number of vital body functions. Research inthis area has identified variants of the alpha subunits that result inmajor changes in channel function and activities, which can ultimatelylead to major pathophysiological conditions. The members of this familyof proteins are denoted NaV1.1 to NaV1.9.

NaV1.7 is a tetrodotoxin-sensitive voltage-gated sodium channel encodedby the gene SCN9A. Human NaV1.7 was first cloned from neuroendocrinecells (Klugbauer, N., et al., 1995 EMBO J., 14 (6): 1084-90.) and ratNaV1.7 was cloned from a pheochromocytoma PC12 cell line (Toledo-Aral,J. J., et al., Proc. Natl. Acad. Sci. USA (1997), 94:1527-1532) and fromrat dorsal root ganglia (Sangameswaran, L., et al., (1997), J. Biol.Chem., 272 (23): 14805-9). NaV1.7 is expressed primarily in theperipheral nervous system, especially nocieptors and olfactory neuronsand sympathetic neurons. The inhibition, or blocking, of NaV1.7 has beenshown to result in analgesic activity. Knockout of NaV1.7 expression ina subset of sensory neurons that are predominantly nociceptive resultsin resistance to inflammatory pain (Nassar, et al., op. cit.). Likewise,loss of function mutations in humans results in congenital indifferenceto pain (CIP), in which the individuals are resistant to bothinflammatory and neuropathic pain (Cox, J. J., et al., Nature (2006);444:894-898; Goldberg, Y. P., et al., Clin. Genet. (2007); 71:311-319).Conversely, gain of function mutations in NaV1.7 have been establishedin two human heritable pain conditions, primary erythromelalgia andfamilial rectal pain, (Yang, Y., et al., J. Med. Genet. (2004), 41(3):171-4). In addition, a single nucleotide polymorphism (R1150W) that hasvery subtle effects on the time- and voltage-dependence of channelgating has large effects on pain perception (Estacion, M., et al., 2009.Ann Neurol 66: 862-6; Reimann, F., et al., Proc Natl Acad Sci USA(2010), 107: 5148-53). About 10% of the patients with a variety of painconditions have the allele conferring greater sensitivity to pain andthus might be more likely to respond to block of NaV1.7. Because NaV1.7is expressed in both sensory and sympathetic neurons, one might expectthat enhanced pain perception would be accompanied by cardiovascularabnormalities such as hypertension, but no correlation has beenreported. Thus, both the CIP mutations and SNP analysis suggest thathuman pain responses are more sensitive to changes in NaV1.7 currentsthan are perturbations of autonomic function.

Sodium channel blockers have been shown to be useful in the treatment ofpain, (see, e.g., Wood, J. N., et al., J. Neurobiol. (2004), 61(1),55-71. Genetic and functional studies have provided evidence to supportthat activity of NaV1.7 as a major contributor to pain signalling inmammals. (See Hajj, et al. Nature Reviews Neuroscience; 2013, vol 14,49-62; and Lee, et al. Cell; 2014, vol 157; 1-12). Presently, there area limited number of effective sodium channel blockers for the treatmentof pain with a minimum of adverse side effects which are currently inthe clinic. Thus there remains a need for selective voltage-gated sodiumchannel modulators (e.g., modulators of NaV1.7) that can provide agreater therapeutic index for treatment.

SUMMARY

In one aspect the present invention provides novel compounds havingsodium channel blocking activity that are useful for the treatment ofpain.

In a first embodiment (Embodiment 1; abbreviated as “E1”) the inventionprovides a compound of the invention, which is a compound of formula(I):

or a pharmaceutically acceptable salt thereof, wherein,

A is a 6-12 membered aryl ring that is optionally substituted with oneor more groups R^(A) independently selected from the group consisting ofF, Cl, Br, I, —CN, —C(═O)OR^(A1), —SO₂R^(A1), —OR^(A1), —(X^(RA))-(3-15membered carbocyclyl), —(X^(RA))-(6-12 membered aryl), —(X^(RA))-(5-12membered heteroaryl), and —R^(A2), wherein said 3-15 memberedcarbocyclyl, 6-12 membered aryl, and 5-12 membered heteroaryl of R^(A)is optionally substituted with from 1 to 5 substitutents R^(a)independently selected from the group consisting of F, Cl, Br, I, C₁₋₄alkyl, halo(C₁₋₄alkyl), amino, (C₁₋₄ alkyl)amino, di(C₁₋₄alkyl)amino,halo(C₁₋₄alkoxy), and —C(O)N(R)₂; R^(AI) is selected from the groupconsisting of hydrogen, C₁₋₈ alkyl, C₂₋₈ alkenyl, halo(C₁₋₈alkyl), C₃₋₈cycloalkyl, phenyl and benzyl; R^(A2) is selected from the groupconsisting of C₁₋₈ alkyl that is optionally substituted with one or moresubstituents independently selected from oxo (═O), fluoro, hydroxy,amino, (C₁₋₄ alkyl)amino and di(C₁₋₄alkyl)amino; X^(RA) is selected fromthe group consisting of absent, —C(═O)—, and C₁₋₄ alkylene; wherein anyC₁₋₄ alkylene, of X^(RA) is optionally substituted with 1 to 3substituents independently selected from the group consisting of C₁₋₄alkyl, halo(C₁₋₄alkyl), and phenyl that is optionally substituted with 1to 5 substitutents independently selected from, F, Cl, Br, I, —NH₂, —OH,—CN, —NO₂, C₁₋₄ alkyl, halo(C₁₋₄alkyl), C₁₋₄ alkoxy, halo(C₁₋₄alkoxy),(C₁₋₄ alkyl)amino and di(C₁₋₄alkyl)amino;

B is a 5-12 membered heteroaryl or a 6-12 membered aryl ring, whereinring B is optionally substituted with one or more groups R^(B)independently selected from the group consisting of F, Cl, Br, I, —CN,—C(═O)OR^(B1), —SO₂R^(B1), —OR^(B1), —(X^(RB))-(3-15 memberedcarbocyclyl), —(X^(RB))-(6-12 membered aryl), —(X^(RB))-(5-12 memberedheteroaryl), and —R^(B2), wherein said 3-15 membered carbocyclyl, 6-12membered aryl, and 5-12 membered heteroaryl of R^(B) is optionallysubstituted with from 1 to 5 substitutents R^(b) independently selectedfrom the group consisting of F, Cl, Br, I, C₁₋₄ alkyl, halo(C₁₋₄alkyl),amino, (C₁₋₄ alkyl)amino, di(C₁₋₄alkyl)amino, halo(C₁₋₄alkoxy), and—C(O)N(R)₂; RBI is selected from the group consisting of hydrogen, C₁₋₈alkyl, C₂₋₈ alkenyl, halo(C₁₋₈alkyl), C₃₋₈ cycloalkyl, phenyl andbenzyl; R^(B2) is selected from the group consisting of C₁₋₈ alkyl thatis optionally substituted with one or more substituents independentlyselected from oxo (═O), fluoro, hydroxy, amino, (C₁₋₄ alkyl)amino anddi(C₁₋₄alkyl)amino; X^(RB) is selected from the group consisting ofabsent, —C(═O)—, and C₁₋₄ alkylene; wherein any C₁₋₄ alkylene, of X^(RB)is optionally substituted with 1 to 3 substituents independentlyselected from the group consisting of C₁₋₄ alkyl, halo(C₁₋₄alkyl), andphenyl that is optionally substituted with 1 to 5 substitutentsindependently selected from, F, Cl, Br, I, —NH₂, —OH, —CN, —NO₂, C₁₋₄alkyl, halo(C₁₋₄alkyl), C₁₋₄ alkoxy, halo(C₁₋₄alkoxy), (C₁₋₄ alkyl)aminoand di(C₁₋₄ alkyl)amino;

ring C is a 5-, 6-, or 7-membered heterocyclyl, which 5-, 6-, or7-membered heterocyclyl is optionally substituted with one or moregroups R^(c) independently selected from the group consisting ofC₁₋₄alkyl, halo, oxo (═O), thioxo (═S), and halo(C₁₋₄ alkyl);

R³, R⁴, and R⁵ are each independently selected from the group consistingof H, F, C₁₋₄ alkyl, C₃₋₈ cycloalkyl, halo(C₁₋₈alkyl) and C₁₋₈ alkoxy;and

R⁶ is selected from the group consisting of H, C₁₋₈ alkyl, C₃₋₈cycloalkyl, aryl, and aryl(C₁₋₄ alkyl); and R⁷ is selected from thegroup consisting of C₁₋₈ alkyl, 3-15 membered heterocyclyl, 5-12membered heteroaryl, —C(O)N(R^(d))₂ and —C(═NCN)N(R^(d))₂; or R⁶ and R⁷,together with the nitrogen to which they are attached, form a 3-15membered heterocyclyl or 5-12 membered heteroaryl; wherein anyC₁₋₈alkyl, C₃₋₈cycloalkyl, aryl, aralkyl, 3-15 membered heterocyclyl,and 5-12 membered heteroaryl is optionally substituted with one or moregroups R^(e) that are independently selected from the group consistingof F, Cl, Br, I, —NH₂, —OH, —CN, —NO₂, C₁₋₄ alkyl, halo(C₁₋₄alkyl), C₁₋₄alkoxy, halo(C₁₋₄alkoxy), (C₁₋₄ alkyl)amino and di(C₁₋₄alkyl)amino; eachR^(d) is independently selected from the group consisting of hydrogen,C₁₋₈ alkyl, C₃₋₈ cycloalkyl, phenyl and benzyl.

In a second embodiment (Embodiment 2; abbreviated as “E2”) the inventionprovides for a compound of formula I which is a compound of formula Ia:

wherein, X is CH₂, O, S, SO, or SO₂; Y is H₂ or O; and Z is CH₂ or O;provided that when Z is O, X is CH₂.

In another embodiment (Embodiment 3; abbreviated as “E3”) the inventionprovides for a compound of formula I which is a compound of formula Ib:

wherein, X is CH₂, O, S, SO, or SO₂; Y is H₂, or O; Z is CH₂ or O;provided that when Z is O, X is CH₂; and p is 1, 2, 3, or 4, or apharmaceutically acceptable salt thereof.

In another embodiment (Embodiment 4; abbreviated as “E4”) the inventionprovides for a compound of formula I which is a compound of formula Ic:

wherein, X is CH₂, O, S, SO, or SO₂; Y is H₂, or O; Z is CH₂ or O;provided that when Z is O, X is CH₂; and p is 1, 2, 3, or 4, or apharmaceutically acceptable salt thereof.

In another embodiment (Embodiment 5; abbreviated as “E5”) the inventionprovides for a compound of formula I which is a compound of formula Id:

wherein, X is CH₂, O, S, SO, or SO₂; Y is H₂, or O; Z is CH₂ or O;provided that when Z is O, X is CH₂; p is 1, 2, 3, or 4; and o is 1, 2,3, 4, or 5; or a pharmaceutically acceptable salt thereof.

In another embodiment (Embodiment 6; abbreviated as “E6”) the inventionprovides for a compound of formula I which is a compound of formula Ie:

wherein, Y is H₂, or O; p is 1, 2, 3, or 4; and o is 1, 2, 3, 4, or 5;or a pharmaceutically acceptable salt thereof.

In another embodiment (Embodiment 7; abbreviated as “E7”) the inventionprovides for a compound of formula I which is a compound of formula If:

wherein, X is CH₂, O, S, SO, or SO₂; Y is H₂, or O; Z is CH₂ or O;provided that when Z is O, X is CH₂; and o is 1, 2, 3, 4, or 5; or apharmaceutically acceptable salt thereof.

In another embodiment (Embodiment 8; abbreviated as “E8”) the inventionprovides for a compound of formula I which is a compound of formula Ig:

wherein, Y is H₂, or O; and o is 1, 2, 3, 4, or 5; or a pharmaceuticallyacceptable salt thereof.

Further embodiments of the compounds of the invention are describedbelow.

E9. A compound of E2, E3, E4, E5, or E7, or a pharmaceuticallyacceptable salt thereof, wherein X is O, S, SO, or SO₂; Y is H₂; and Zis CH₂.

E10. A compound of E2, E3, E4, E5, or E7, or a pharmaceuticallyacceptable salt thereof, wherein X is O, S, SO, or SO₂; Y is O; and Z isCH₂.

E11. A compound of E6 or E8, or a pharmaceutically acceptable saltthereof, wherein Y is H₂.

E12. A compound of E6 or E8, or a pharmaceutically acceptable saltthereof, wherein Y is O.

E13. A compound of E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, or E12,or a pharmaceutically acceptable salt thereof, wherein R³ is H.

E14. A compound of E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, or E12,or a pharmaceutically acceptable salt thereof, wherein R⁴ is H.

E15. A compound of E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, or E12,or a pharmaceutically acceptable salt thereof, wherein R⁵ is H.

E16. A compound of E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, or E12,or a pharmaceutically acceptable salt thereof, wherein R³, R⁴, and R⁵are each independently selected from the group consisting of H, F, andC₁₋₄ alkyl.

E17. A compound of E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, or E12,or a pharmaceutically acceptable salt thereof, wherein R³, R⁴, and R⁵are each H.

E18. A compound of E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12,E13, E14, E15, E16 or E17, or a pharmaceutically acceptable saltthereof, wherein —NR⁶R⁷ is:

E19. A compound of E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12,E13, E14, E15, E16 or E17, or a pharmaceutically acceptable saltthereof, wherein —NR⁶R⁷ is:

E20. A compound of E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12,E13, E14, E15, E16 or E17, or a pharmaceutically acceptable saltthereof, wherein R⁶ is H and R⁷ is 5-6 membered heteroaryl.

E21. A compound of E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12,E13, E14, E15, E16, E17, E18, E19, or E20, or a pharmaceuticallyacceptable salt thereof, wherein each R^(A) is independently selectedfrom the group consisting of, F, Cl, Br, I, —OR^(A1), and —R^(A2),wherein R^(A1) is selected from the group consisting of C₁₋₈ alkyl andhalo(C₁₋₈alkyl), and wherein R^(A2) is C₁₋₈ alkyl that is optionallysubstituted with one or more substituents independently selected fromthe group consisting of fluoro and chloro.

E22. A compound of E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12,E13, E14, E15, E16, E17, E18, E19, E20, or E21, or a pharmaceuticallyacceptable salt thereof, wherein each R^(B) is independently selectedfrom the group consisting of, F, Cl, Br, I, —CN, and —OR^(A1), whereinR^(A1) is selected from the group consisting of C₁₋₈ alkyl andhalo(C₁₋₈alkyl).

E23. A compound of E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12,E13, E14, E15, E16, E17, E18, E19, E20, E21 or E22, or apharmaceutically acceptable salt thereof, wherein each R^(A) isindependently selected from the group consisting of, F, Cl,trifluoromethoxy, CF₂, and CF₃.

E24. A compound of E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12,E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, or E23, or apharmaceutically acceptable salt thereof, wherein each R^(B) isindependently selected from the group consisting of, F, —CN, andmethoxy.

In another aspect the present invention provides for a pharmaceuticalcomposition comprising a compound as described herein or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable excipient.

In another aspect the present invention provides for a method oftreating a disease or condition in a mammal selected from the groupconsisting of pain, depression, cardiovascular diseases, respiratorydiseases, and psychiatric diseases, and combinations thereof, whereinthe method comprises administering to the mammal in need thereof atherapeutically effective amount of a compound as described herein, or apharmaceutically acceptable salt thereof. In another aspect of thepresent invention said disease or condition is selected from the groupconsisting of neuropathic pain, inflammatory pain, visceral pain, cancerpain, chemotherapy pain, trauma pain, surgical pain, post-surgical pain,childbirth pain, labor pain, neurogenic bladder, ulcerative colitis,chronic pain, persistent pain, peripherally mediated pain, centrallymediated pain, chronic headache, migraine headache, sinus headache,tension headache, phantom limb pain, dental pain, peripheral nerveinjury or a combination thereof. In another aspect of the presentinvention said disease or condition is selected from the groupconsisting of pain associated with HIV, HIV treatment inducedneuropathy, trigeminal neuralgia, post-herpetic neuralgia, eudynia, heatsensitivity, tosarcoidosis, irritable bowel syndrome, Crohns disease,pain associated with multiple sclerosis (MS), amyotrophic lateralsclerosis (ALS), diabetic neuropathy, peripheral neuropathy, arthritis,rheumatoid arthritis, osteoarthritis, atherosclerosis, paroxysmaldystonia, myasthenia syndromes, myotonia, malignant hyperthermia, cysticfibrosis, pseudoaldosteronism, rhabdomyolysis, hypothyroidism, bipolardepression, anxiety, schizophrenia, sodium channel toxin relatedillnesses, familial erythromelalgia, primary erythromelalgia, familialrectal pain, cancer, epilepsy, partial and general tonic seizures,restless leg syndrome, arrhythmias, fibromyalgia, neuroprotection underischaemic conditions cause by stroke or neural trauma, tach-arrhythmias,atrial fibrillation and ventricular fibrillation.

In another aspect the present invention provides for a method oftreating pain in a mammal by the inhibition of ion flux through avoltage-dependent sodium channel in the mammal, wherein the methodcomprises administering to the mammal in need thereof a therapeuticallyeffective amount of a compound as described herein, or apharmaceutically acceptable salt thereof.

In another aspect the present invention provides for a method ofdecreasing ion flux through a voltage-dependent sodium channel in a cellin a mammal, wherein the method comprises contacting the cell with acompound as described herein, or a pharmaceutically acceptable saltthereof.

In another aspect the present invention provides for a method oftreating pruritus in a mammal, wherein the method comprisesadministering to the mammal in need thereof a therapeutically effectiveamount of a compound of the invention, or a pharmaceutically acceptablesalt thereof.

In another aspect the present invention provides for a method oftreating cancer in a mammal, wherein the method comprises administeringto the mammal in need thereof a therapeutically effective amount acompound as described herein, or a pharmaceutically acceptable saltthereof.

In another aspect the present invention provides for a method oftreating, but not preventing, pain in a mammal, wherein the methodcomprises administering to the mammal in need thereof a therapeuticallyeffective amount of a compound as described herein, or apharmaceutically acceptable salt thereof. In another aspect of thepresent invention the pain is selected from the group consisting ofneuropathic pain, inflammatory pain, visceral pain, cancer pain,chemotherapy pain, trauma pain, surgical pain, post-surgical pain,childbirth pain, labor pain, neurogenic bladder, ulcerative colitis,chronic pain, persistent pain, peripherally mediated pain, centrallymediated pain, chronic headache, migraine headache, sinus headache,tension headache, phantom limb pain, dental pain, peripheral nerveinjury or a combination thereof. In another aspect the present inventionthe pain is associated with a disease or condition selected from thegroup consisting of HIV, HIV treatment induced neuropathy, trigeminalneuralgia, post-herpetic neuralgia, eudynia, heat sensitivity,tosarcoidosis, irritable bowel syndrome, Crohns disease, pain associatedwith multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS),diabetic neuropathy, peripheral neuropathy, arthritis, rheumatoidarthritis, osteoarthritis, atherosclerosis, paroxysmal dystonia,myasthenia syndromes, myotonia, malignant hyperthermia, cystic fibrosis,pseudoaldosteronism, rhabdomyolysis, hypothyroidism, bipolar depression,anxiety, schizophrenia, sodium channel toxin related illnesses, familialerythromelalgia, primary erythromelalgia, familial rectal pain, cancer,epilepsy, partial and general tonic seizures, restless leg syndrome,arrhythmias, fibromyalgia, neuroprotection under ischaemic conditionscause by stroke or neural trauma, tach-arrhythmias, atrial fibrillationand ventricular fibrillation.

In another aspect the present invention provides for a method for thetreatment or prophylaxis of pain, depression, cardiovascular disease,respiratory disease, or psychiatric disease, or a combinations thereof,in an animal which method comprises administering an effective amount ofa compound of as described herein, or a pharmaceutically acceptable saltthereof.

In another aspect the present invention provides for a compound asdescribed herein, or a pharmaceutically acceptable salt thereof for theuse as a medicament for the treatment of diseases and disorders selectedfrom the group consisting of pain, depression, cardiovascular diseases,respiratory diseases, and psychiatric diseases, or a combinationthereof.

In another aspect the present invention provides for the use of acompound as described herein, or a pharmaceutically acceptable saltthereof for the manufacture of a medicament for the treatment ofdiseases and disorders selected from the group consisting of pain,depression, cardiovascular diseases, respiratory diseases, andpsychiatric diseases, or a combination thereof.

In another aspect the present invention provides for the invention asdescribed herein.

DETAILED DESCRIPTION Definitions

As used herein, the term “alkyl”, by itself or as part of anothersubstituent, means, unless otherwise stated, a straight or branchedchain hydrocarbon radical, having the number of carbon atoms designated(i.e., C₁₋₈ means one to eight carbons). Examples of alkyl groupsinclude methyl, ethyl, n-propyl, iso-propyl, n-butyl, t-butyl,iso-butyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and thelike.

The terms “alkoxy,” “alkylamino” and “alkylthio”, are used in theirconventional sense, and refer to those alkyl groups attached to theremainder of the molecule via an oxygen atom (“oxy”), an amino group(“amino”) or thio group. Additionally, for dialkylamino groups, thealkyl portions can be the same or different.

The terms “halo” by itself or as part of another substituent, mean,unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.

The term “halo(C_(1-x)alkyl)” refers to an alkyl that has 1-x carbonatoms and that is substituted with one or more (e.g. 1, 2, 3, 4, 5, or6) halo groups. For example the term includes an alkyl group having 1-6carbon atoms that is substituted with one or more halo groups.Non-limiting examples of the term halo(C₁-C₆alkyl) include fluoromethyl,difluoromethyl, trifluoromethyl, chloromethyl, and 2,2,2-trifluoroethyl.

The term “halo(C_(1-x)alkoxy)” refers to an alkoxy group that has 1-xcarbon atoms and that is substituted with one or more (e.g. 1, 2, 3, 4,5, or 6) halo groups. For example the term includes an alkoxy grouphaving 1-6 carbon atoms that is substituted with one or more halogroups. Non-limiting examples of the term halo(C₁-C₆alkyl) includefluoromethoxy, difluoromethoxy, trifluoromethoxy, chloromethoxy, and2,2,2-trifluoroethoxy.

The term “aryl” as used herein refers to a single all carbon aromaticring or a multiple condensed all carbon ring system wherein at least oneof the rings is aromatic. For example, in certain embodiments, an arylgroup has 6 to 12 carbon atoms. Aryl includes a phenyl radical. Arylalso includes multiple condensed ring systems (e.g., ring systemscomprising 2, 3 or 4 rings) having about 9 to 12 carbon atoms in whichat least one ring is aromatic and wherein the other rings may bearomatic or not aromatic (i.e., carbocycle). Such multiple condensedring systems are optionally substituted with one or more (e.g., 1, 2 or3) oxo groups on any carbocycle portion of the multiple condensed ringsystem. The rings of the multiple condensed ring system can be connectedto each other via fused, spiro and bridged bonds when allowed by valencyrequirements. It is to be understood that the point of attachment of amultiple condensed ring system, as defined above, can be at any positionof the ring system including an aromatic or a carbocycle portion of thering. Non-limiting examples of aryl groups include, but are not limitedto, phenyl, indenyl, naphthyl, 1, 2, 3, 4-tetrahydronaphthyl, and thelike.

The term “carbocycle” or “carbocyclyl” refers to a single saturated(i.e., cycloalkyl) or a single partially unsaturated (e.g.,cycloalkenyl, cycloalkadienyl, etc.) all carbon ring having 3 to 7carbon atoms (i.e., (C₃-C₇)carbocycle). The term “carbocycle” or“carbocyclyl” also includes multiple condensed, saturated and partiallyunsaturated all carbon ring systems (e.g., ring systems comprising 2, 3or 4 carbocyclic rings). Accordingly, carbocycle includes multicycliccarbocyles such as a bicyclic carbocycles (e.g., bicyclic carbocycleshaving about 6 to 15 or 6 to 12 carbon atoms such asbicyclo[3.1.0]hexane and bicyclo[2.1.1]hexane), and polycycliccarbocycles (e.g tricyclic and tetracyclic carbocycles with up to about15 carbon atoms). The rings of the multiple condensed ring system can beconnected to each other via fused, spiro and bridged bonds when allowedby valency requirements. For example, multicyclic carbocyles can beconnected to each other via a single carbon atom to form a spiroconnection (e.g., spiropentane, spiro[4,5]decane, etc), via two adjacentcarbon atoms to form a fused connection (e.g., carbocycles such asdecahydronaphthalene, norsabinane, norcarane) or via two non-adjacentcarbon atoms to form a bridged connection (e.g., norbornane,bicyclo[2.2.2]octane, etc). The “carbocycle” or “carbocyclyl” can alsobe optionally substituted with one or more (e.g., 1, 2 or 3) oxo groups.In one embodiment the term carbocycle includes a C₃₋₁₅ carbocycle. Inone embodiment the term carbocycle includes a C₃₋₁₂ carbocycle. In oneembodiment the term carbocycle includes a C₃₋₈ carbocycle. In oneembodiment the term carbocycle includes a C₃₋₆ carbocycle. In oneembodiment the term carbocycle includes a C₃₋₅ carbocycle. Non-limitingexamples of carbocycles include cyclopropyl, cyclobutyl, cyclopentyl,1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl,1-cyclohex-1-enyl, 1-cyclohex-2-enyl, bicyclo[2.2.1]heptane, pinane,adamantane, norborene, spirocyclic C₅₋₁₂ alkane, and 1-cyclohex-3-enyl.In one embodiment the term “cycloalkyl” refers to a single saturated allcarbon ring having 3 to 8 carbon atoms. Non-limiting examples ofcarbocycles include cyclopropyl, cyclobutyl, cyclopentyl, andcyclohexyl.

The term “heteroaryl” as used herein refers to a single aromatic ringthat has at least one atom other than carbon in the ring, wherein theatom is selected from the group consisting of oxygen, nitrogen andsulfur; “heteroaryl” also includes multiple condensed ring systems thathave at least one such aromatic ring, which multiple condensed ringsystems are further described below. Thus, “heteroaryl” includes singlearomatic rings of from about 1 to 6 carbon atoms and about 1-4heteroatoms selected from the group consisting of oxygen, nitrogen andsulfur. The sulfur and nitrogen atoms may also be present in an oxidizedform provided the ring is aromatic. Exemplary heteroaryl ring systemsinclude but are not limited to pyridyl, pyrimidinyl, oxazolyl or furyl.“Heteroaryl” also includes multiple condensed ring systems (e.g., ringsystems comprising 2, 3 or 4 rings) wherein a heteroaryl group, asdefined above, is condensed with one or more rings selected fromheteroaryls (to form for example a naphthyridinyl such as1,8-naphthyridinyl), heterocycles, (to form for example a 1, 2, 3,4-tetrahydronaphthyridinyl such as1,2,3,4-tetrahydro-1,8-naphthyridinyl), carbocycles (to form for example5,6,7,8-tetrahydroquinolyl) and aryls (to form for example indazolyl) toform the multiple condensed ring system. Thus, a heteroaryl (a singlearomatic ring or multiple condensed ring system) has about 1-20 carbonatoms and about 1-6 heteroatoms within the heteroaryl ring. A heteroaryl(a single aromatic ring or multiple condensed ring system) can also haveabout 5 to 12 or about 5 to 10 members within the heteroaryl ring.Multiple condensed ring systems may be optionally substituted with oneor more (e.g., 1, 2, 3 or 4) oxo groups on the carbocycle or heterocycleportions of the condensed ring. The rings of a multiple condensed ringsystem can be connected to each other via fused, spiro and bridged bondswhen allowed by valency requirements. It is to be understood that theindividual rings of the multiple condensed ring system may be connectedin any order relative to one another. It is also to be understood thatthe point of attachment of a multiple condensed ring system (as definedabove for a heteroaryl) can be at any position of the multiple condensedring system including a heteroaryl, heterocycle, aryl or carbocycleportion of the multiple condensed ring system. It is also to beunderstood that the point of attachment for a heteroaryl or heteroarylmultiple condensed ring system can be at any suitable atom of theheteroaryl or heteroaryl multiple condensed ring system including acarbon atom and a heteroatom (e.g., a nitrogen). Exemplary heteroarylsinclude but are not limited to pyridyl, pyrrolyl, pyrazinyl,pyrimidinyl, pyridazinyl, pyrazolyl, thienyl, indolyl, imidazolyl,oxazolyl, isoxazolyl, thiazolyl, furyl, oxadiazolyl, thiadiazolyl,quinolyl, isoquinolyl, benzothiazolyl, benzoxazolyl, indazolyl,quinoxalyl, quinazolyl, 5,6,7,8-tetrahydroisoquinolinyl benzofuranyl,benzimidazolyl, thianaphthenyl, pyrrolo[2,3-b]pyridinyl,quinazolinyl-4(3H)-one, triazolyl, 4,5,6,7-tetrahydro-1H-indazole and3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclo-penta[1,2-c]pyrazole. Inone embodiment the term “heteroaryl” refers to a single aromatic ringcontaining at least one heteroatom. For example, the term includes5-membered and 6-membered monocyclic aromatic rings that include one ormore heteroatoms. Non-limiting examples of heteroaryl include but arenot limited to pyridyl, furyl, thiazole, pyrimidine, oxazole, andthiadiazole.

The term “heterocyclyl” or “heterocycle” as used herein refers to asingle saturated or partially unsaturated ring that has at least oneatom other than carbon in the ring, wherein the atom is selected fromthe group consisting of oxygen, nitrogen and sulfur; the term alsoincludes multiple condensed ring systems that have at least one suchsaturated or partially unsaturated ring, which multiple condensed ringsystems are further described below. Thus, the term includes singlesaturated or partially unsaturated rings (e.g., 3, 4, 5, 6 or 7-memberedrings) from about 1 to 6 carbon atoms and from about 1 to 3 heteroatomsselected from the group consisting of oxygen, nitrogen and sulfur in thering. The ring may be substituted with one or more (e.g., 1, 2 or 3) oxogroups and the sulfur and nitrogen atoms may also be present in theiroxidized forms. Exemplary heterocycles include but are not limited toazetidinyl, tetrahydrofuranyl and piperidinyl. The term “heterocycle”also includes multiple condensed ring systems (e.g., ring systemscomprising 2, 3 or 4 rings) wherein a single heterocycle ring (asdefined above) can be condensed with one or more groups selected fromheterocycles (to form for example a 1,8-decahydronapthyridinyl),carbocycles (to form for example a decahydroquinolyl) and aryls to formthe multiple condensed ring system. Thus, a heterocycle (a singlesaturated or single partially unsaturated ring or multiple condensedring system) has about 2-20 carbon atoms and 1-6 heteroatoms within theheterocycle ring. Such multiple condensed ring systems may be optionallysubstituted with one or more (e.g., 1, 2, 3 or 4) oxo groups on thecarbocycle or heterocycle portions of the multiple condensed ring. Therings of the multiple condensed ring system can be connected to eachother via fused, spiro and bridged bonds when allowed by valencyrequirements. It is to be understood that the individual rings of themultiple condensed ring system may be connected in any order relative toone another. Accordingly, a heterocycle (a single saturated or singlepartially unsaturated ring or multiple condensed ring system) has about3-20 atoms including about 1-6 heteroatoms within the heterocycle ringsystem. It is also to be understood that the point of attachment of amultiple condensed ring system (as defined above for a heterocycle) canbe at any position of the multiple condensed ring system including aheterocycle, aryl and carbocycle portion of the ring. It is also to beunderstood that the point of attachment for a heterocycle or heterocyclemultiple condensed ring system can be at any suitable atom of theheterocycle or heterocycle multiple condensed ring system including acarbon atom and a heteroatom (e.g., a nitrogen). In one embodiment theterm heterocycle includes a C₂₋₂₀ heterocycle. In one embodiment theterm heterocycle includes a C₂₋₇ heterocycle. In one embodiment the termheterocycle includes a C₂₋₅ heterocycle. In one embodiment the termheterocycle includes a C₂₋₄ heterocycle. Exemplary heterocycles include,but are not limited to aziridinyl, azetidinyl, pyrrolidinyl,piperidinyl, homopiperidinyl, morpholinyl, thiomorpholinyl, piperazinyl,tetrahydrofuranyl, dihydrooxazolyl, tetrahydropyranyl,tetrahydrothiopyranyl, 1,2,3,4-tetrahydroquinolyl, benzoxazinyl,dihydrooxazolyl, chromanyl, 1,2-dihydropyridinyl,2,3-dihydrobenzofuranyl, 1,3-benzodioxolyl, 1,4-benzodioxanyl,spiro[cyclopropane-1,1′-isoindolinyl]-3′-one, isoindolinyl-1-one,2-oxa-6-azaspiro[3.3]heptanyl, imidazolidin-2-one N-methylpiperidine,imidazolidine, pyrazolidine, butyrolactam, valerolactam,imidazolidinone, hydantoin, dioxolane, phthalimide, 1,4-dioxane,thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, pyran,3-pyrroline, thiopyran, pyrone, tetrhydrothiophene, quinuclidine,tropane, 2-azaspiro[3.3]heptane, (1R,5S)-3-azabicyclo[3.2.1]octane,(1s,4s)-2-azabicyclo[2.2.2]octane,(1R,4R)-2-oxa-5-azabicyclo[2.2.2]octane and pyrrolidin-2-one. In oneembodiment the term “heterocycle” refers to a monocyclic, saturated orpartially unsaturated, 3-8 membered ring having at least one heteroatom.For example, the term includes a monocyclic, saturated or partiallyunsaturated, 4, 5, 6, or 7 membered ring having at least one heteroatom.Non-limiting examples of heterocycle include aziridine, azetidine,pyrrolidine, piperidine, piperidine, piperazine, oxirane, morpholine,and thiomorpholine. The term “9- or 10-membered heterobicycle” as usedherein refers to a partially unsaturated or aromatic fused bicyclic ringsystem having at least one heteroatom. For example, the term 9- or10-membered heterobicycle includes a bicyclic ring system having a benzoring fused to a 5-membered or 6-membered saturated, partiallyunsaturated, or aromatic ring that contains one or more heteroatoms.

As used herein, the term “heteroatom” is meant to include oxygen (O),nitrogen (N), sulfur (S) and silicon (Si). The nitrogen and sulfur canbe in an oxidized form when feasible.

As used herein, the term “chiral” refers to molecules which have theproperty of non-superimposability of the mirror image partner, while theterm “achiral” refers to molecules which are superimposable on theirmirror image partner.

As used herein, the term “stereoisomers” refers to compounds which haveidentical chemical constitution, but differ with regard to thearrangement of the atoms or groups in space.

As used herein a wavy line “

” that intersects a bond in a chemical structure indicates the point ofattachment of the bond that the wavy bond intersects in the chemicalstructure to the remainder of a molecule.

“Diastereomer” refers to a stereoisomer with two or more centers ofchirality and whose molecules are not mirror images of one another.Diastereomers have different physical properties, e.g. melting points,boiling points, spectral properties, and reactivities. Mixtures ofdiastereomers can separate under high resolution analytical proceduressuch as electrophoresis and chromatography.

“Enantiomers” refer to two stereoisomers of a compound which arenon-superimposable mirror images of one another.

Stereochemical definitions and conventions used herein generally followS. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984)McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S.,“Stereochemistry of Organic Compounds”, John Wiley & Sons, Inc., NewYork, 1994. The compounds of the invention can contain asymmetric orchiral centers, and therefore exist in different stereoisomeric forms.It is intended that all stereoisomeric forms of the compounds of theinvention, including but not limited to, diastereomers, enantiomers andatropisomers, as well as mixtures thereof such as racemic mixtures, formpart of the present invention. Many organic compounds exist in opticallyactive forms, i.e., they have the ability to rotate the plane ofplane-polarized light. In describing an optically active compound, theprefixes D and L, or R and S, are used to denote the absoluteconfiguration of the molecule about its chiral center(s). The prefixes dand 1 or (+) and (−) are employed to designate the sign of rotation ofplane-polarized light by the compound, with (−) or 1 meaning that thecompound is levorotatory. A compound prefixed with (+) or d isdextrorotatory. For a given chemical structure, these stereoisomers areidentical except that they are mirror images of one another. A specificstereoisomer can also be referred to as an enantiomer, and a mixture ofsuch isomers is often called an enantiomeric mixture. A 50:50 mixture ofenantiomers is referred to as a racemic mixture or a racemate, which canoccur where there has been no stereoselection or stereospecificity in achemical reaction or process. The terms “racemic mixture” and “racemate”refer to an equimolar mixture of two enantiomeric species, devoid ofoptical activity.

When a bond in a compound formula herein is drawn in anon-stereochemical manner (e.g. flat), the atom to which the bond isattached includes all stereochemical possibilities. When a bond in acompound formula herein is drawn in a defined stereochemical manner(e.g. bold, bold-wedge, dashed or dashed-wedge), it is to be understoodthat the atom to which the stereochemical bond is attached is enrichedin the absolute stereoisomer depicted unless otherwise noted. In oneembodiment, the compound may be at least 51% the absolute stereoisomerdepicted. In another embodiment, the compound may be at least 80% theabsolute stereoisomer depicted. In another embodiment, the compound maybe at least 90% the absolute stereoisomer depicted. In anotherembodiment, the compound may be at least 95% the absolute stereoisomerdepicted. In another embodiment, the compound may be at least 97% theabsolute stereoisomer depicted. In another embodiment, the compound maybe at least 98% the absolute stereoisomer depicted. In anotherembodiment, the compound may be at least 99% the absolute stereoisomerdepicted.

As used herein, the term “tautomer” or “tautomeric form” refers tostructural isomers of different energies which are interconvertible viaa low energy barrier. For example, proton tautomers (also known asprototropic tautomers) include interconversions via migration of aproton, such as keto-enol and imine-enamine isomerizations. Valencetautomers include interconversions by reorganization of some of thebonding electrons.

As used herein, the term “solvate” refers to an association or complexof one or more solvent molecules and a compound of the invention.Examples of solvents that form solvates include, but are not limited to,water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid,and ethanolamine. The term “hydrate” refers to the complex where thesolvent molecule is water.

As used herein, the term “protecting group” refers to a substituent thatis commonly employed to block or protect a particular functional groupon a compound. For example, an “amino-protecting group” is a substituentattached to an amino group that blocks or protects the aminofunctionality in the compound. Suitable amino-protecting groups includeacetyl, trifluoroacetyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBZ)and 9-fluorenylmethylenoxycarbonyl (Fmoc). Similarly, a“hydroxy-protecting group” refers to a substituent of a hydroxy groupthat blocks or protects the hydroxy functionality. Suitable protectinggroups include acetyl and silyl. A “carboxy-protecting group” refers toa substituent of the carboxy group that blocks or protects the carboxyfunctionality. Common carboxy-protecting groups includephenylsulfonylethyl, cyanoethyl, 2-(trimethylsilyl)ethyl,2-(trimethylsilyl)ethoxymethyl, 2-(p-toluenesulfonyl)ethyl,2-(p-nitrophenylsulfenyl)ethyl, 2-(diphenylphosphino)-ethyl, nitroethyland the like. For a general description of protecting groups and theiruse, see P. G. M. Wuts and T. W. Greene, Greene's Protective Groups inOrganic Synthesis 4^(th) edition, Wiley-Interscience, New York, 2006.

As used herein, the term “mammal” includes, but is not limited to,humans, mice, rats, guinea pigs, monkeys, dogs, cats, horses, cows,pigs, and sheep.

As used herein, Y is “H₂” means that Y is two hydrogens, forming CH₂when taken together with the carbon to which Y is attached.

As used herein, the term “pharmaceutically acceptable salts” is meant toinclude salts of the active compounds which are prepared with relativelynontoxic acids or bases, depending on the particular substituents foundon the compounds described herein. When compounds of the presentinvention contain relatively acidic functionalities, base addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of salts derived frompharmaceutically-acceptable inorganic bases include aluminum, ammonium,calcium, copper, ferric, ferrous, lithium, magnesium, manganic,manganous, potassium, sodium, zinc and the like. Salts derived frompharmaceutically-acceptable organic bases include salts of primary,secondary and tertiary amines, including substituted amines, cyclicamines, naturally-occurring amines and the like, such as arginine,betaine, caffeine, choline, N,N′-dibenzylethylenediamine, diethylamine,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperidine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine and the like. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, malonic, benzoic, succinic, suberic,fumaric, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric,tartaric, methanesulfonic, and the like. Also included are salts ofamino acids such as arginate and the like, and salts of organic acidslike glucuronic or galactunoric acids and the like (see, for example,Berge, S. M., et al., “Pharmaceutical Salts”, Journal of PharmaceuticalScience, 1977, 66, 1-19). Certain specific compounds of the presentinvention contain both basic and acidic functionalities that allow thecompounds to be converted into either base or acid addition salts.

The neutral forms of the compounds can be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

In addition to salt forms, the present invention provides compoundswhich are in a prodrug form. As used herein the term “prodrug” refers tothose compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

Prodrugs of the invention include compounds wherein an amino acidresidue, or a polypeptide chain of two or more (e.g., two, three orfour) amino acid residues, is covalently joined through an amide orester bond to a free amino, hydroxy or carboxylic acid group of acompound of the present invention. The amino acid residues include butare not limited to the 20 naturally occurring amino acids commonlydesignated by three letter symbols and also includes phosphoserine,phosphothreonine, phosphotyrosine, 4-hydroxyproline, hydroxylysine,demosine, isodemosine, gamma-carboxyglutamate, hippuric acid,octahydroindole-2-carboxylic acid, statine,1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, penicillamine,ornithine, 3-methylhistidine, norvaline, beta-alanine,gamma-aminobutyric acid, citrulline, homocysteine, homoserine,methyl-alanine, para-benzoylphenylalanine, phenylglycine,propargylglycine, sarcosine, methionine sulfone and tert-butylglycine.

Additional types of prodrugs are also encompassed. For instance, a freecarboxyl group of a compound of the invention can be derivatized as anamide or alkyl ester. As another example, compounds of this inventioncomprising free hydroxy groups can be derivatized as prodrugs byconverting the hydroxy group into a group such as, but not limited to, aphosphate ester, hemisuccinate, dimethylaminoacetate, orphosphoryloxymethyloxycarbonyl group, as outlined in Fleisher, D. etal., (1996) Improved oral drug delivery: solubility limitations overcomeby the use of prodrugs Advanced Drug Delivery Reviews, 19:115.

Carbamate prodrugs of hydroxy and amino groups are also included, as arecarbonate prodrugs, sulfonate esters and sulfate esters of hydroxygroups. Derivatization of hydroxy groups as (acyloxy)methyl and(acyloxy)ethyl ethers, wherein the acyl group can be an alkyl esteroptionally substituted with groups including, but not limited to, ether,amine and carboxylic acid functionalities, or where the acyl group is anamino acid ester as described above, are also encompassed. Prodrugs ofthis type are described in J. Med. Chem., (1996), 39:10. More specificexamples include replacement of the hydrogen atom of the alcohol groupwith a group such as (C₁₋₆)alkanoyloxymethyl,1-((C₁₋₆)alkanoyloxy)ethyl, 1-methyl-1-((C₁₋₆)alkanoyloxy)ethyl,(C₁₋₆)alkoxycarbonyloxymethyl, N—(C₁₋₆)alkoxycarbonylaminomethyl,succinoyl, (C₁₋₆)alkanoyl, alpha-amino(C₁₋₄)alkanoyl, arylacyl andalpha-aminoacyl, or alpha-aminoacyl-alpha-aminoacyl, where eachalpha-aminoacyl group is independently selected from the naturallyoccurring L-amino acids, P(O)(OH)₂, —P(O)(O(C₁₋₆)alkyl)₂ or glycosyl(the radical resulting from the removal of a hydroxyl group of thehemiacetal form of a carbohydrate).

For additional examples of prodrug derivatives, see, for example, a)Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985) and Methodsin Enzymology, Vol. 42, p. 309-396, edited by K. Widder, et al.(Academic Press, 1985); b) A Textbook of Drug Design and Development,edited by Krogsgaard-Larsen and H. Bundgaard, Chapter 5 “Design andApplication of Prodrugs,” by H. Bundgaard p. 113-191 (1991); c) H.Bundgaard, Advanced Drug Delivery Reviews, 8:1-38 (1992); d) H.Bundgaard, et al., Journal of Pharmaceutical Sciences, 77:285 (1988);and e) N. Kakeya, et al., Chem. Pharm. Bull., 32:692 (1984), each ofwhich is specifically incorporated herein by reference.

Additionally, the present invention provides for metabolites ofcompounds of the invention. As used herein, a “metabolite” refers to aproduct produced through metabolism in the body of a specified compoundor salt thereof. Such products can result for example from theoxidation, reduction, hydrolysis, amidation, deamidation,esterification, deesterification, enzymatic cleavage, and the like, ofthe administered compound.

Metabolite products typically are identified by preparing aradiolabelled (e.g., 14C or ³H) isotope of a compound of the invention,administering it parenterally in a detectable dose (e.g., greater thanabout 0.5 mg/kg) to an animal such as rat, mouse, guinea pig, monkey, orto man, allowing sufficient time for metabolism to occur (typicallyabout 30 seconds to 30 hours) and isolating its conversion products fromthe urine, blood or other biological samples. These products are easilyisolated since they are labeled (others are isolated by the use ofantibodies capable of binding epitopes surviving in the metabolite). Themetabolite structures are determined in conventional fashion, e.g., byMS, LC/MS or NMR analysis. In general, analysis of metabolites is donein the same way as conventional drug metabolism studies well known tothose skilled in the art. The metabolite products, so long as they arenot otherwise found in vivo, are useful in diagnostic assays fortherapeutic dosing of the compounds of the invention.

Pharmaceutical Compositions and Administration

In addition to one or more of the compounds provided above (orstereoisomers, geometric isomers, tautomers, solvates, metabolites,isotopes, pharmaceutically acceptable salts, or prodrugs thereof), theinvention also provides for compositions and medicaments comprising acompound of the invention and at least one pharmaceutically acceptablecarrier, diluent or excipient. The compositions of the invention can beused to selectively inhibit NaV1.7 in patients (e.g, humans).

The term “composition,” as used herein, is intended to encompass aproduct comprising the specified ingredients in the specified amounts,as well as any product which results, directly or indirectly, fromcombination of the specified ingredients in the specified amounts. By“pharmaceutically acceptable” it is meant the carrier, diluent orexcipient must be compatible with the other ingredients of theformulation and not deleterious to the recipient thereof.

In one embodiment, the invention provides for pharmaceuticalcompositions (or medicaments) comprising a compound as described herein,and its stereoisomers, geometric isomers, tautomers, solvates,metabolites, isotopes, pharmaceutically acceptable salts, or prodrugsthereof) and a pharmaceutically acceptable carrier, diluent orexcipient. In another embodiment, the invention provides for preparingcompositions (or medicaments) comprising compounds of the invention. Inanother embodiment, the invention provides for administering a compoundof the invention or a and compositions comprising a compound of theinvention to a patient (e.g., a human patient) in need thereof.

Compositions are formulated, dosed, and administered in a fashionconsistent with good medical practice. Factors for consideration in thiscontext include the particular disorder being treated, the particularmammal being treated, the clinical condition of the individual patient,the cause of the disorder, the site of delivery of the agent, the methodof administration, the scheduling of administration, and other factorsknown to medical practitioners. The effective amount of the compound tobe administered will be governed by such considerations, and is theminimum amount necessary to inhibit NaV1.7 activity as required toprevent or treat the undesired disease or disorder, such as for example,pain. For example, such amount may be below the amount that is toxic tonormal cells, or the mammal as a whole.

In one example, the therapeutically effective amount of the compound ofthe invention administered parenterally per dose will be in the range ofabout 0.01-100 mg/kg, alternatively about e.g., 0.1 to 20 mg/kg ofpatient body weight per day, with the typical initial range of compoundused being 0.3 to 15 mg/kg/day. The daily does is, in certainembodiments, given as a single daily dose or in divided doses two to sixtimes a day, or in sustained release form. In the case of a 70 kg adulthuman, the total daily dose will generally be from about 7 mg to about1,400 mg. This dosage regimen may be adjusted to provide the optimaltherapeutic response. The compounds may be administered on a regimen of1 to 4 times per day, preferably once or twice per day.

The compounds of the present invention may be administered in anyconvenient administrative form, e.g., tablets, powders, capsules,solutions, dispersions, suspensions, syrups, sprays, suppositories,gels, emulsions, patches, etc. Such compositions may contain componentsconventional in pharmaceutical preparations, e.g., diluents, carriers,pH modifiers, sweeteners, bulking agents, and further active agents.

The compounds of the invention may be administered by any suitablemeans, including oral, topical (including buccal and sublingual),rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal,intrapulmonary, intradermal, intrathecal and epidural and intranasal,and, if desired for local treatment, intralesional administration.Parenteral infusions include intramuscular, intravenous, intraarterial,intraperitoneal, intracerebral, intraocular, intralesional orsubcutaneous administration.

The compositions comprising compounds as described herein or anembodiment thereof are normally formulated in accordance with standardpharmaceutical practice as a pharmaceutical composition. A typicalformulation is prepared by mixing a compound of the present inventionand a diluent, carrier or excipient. Suitable diluents, carriers andexcipients are well known to those skilled in the art and are describedin detail in, e.g., Ansel, Howard C., et al., Ansel's PharmaceuticalDosage Forms and Drug Delivery Systems. Philadelphia: Lippincott,Williams & Wilkins, 2004; Gennaro, Alfonso R., et al. Remington: TheScience and Practice of Pharmacy. Philadelphia: Lippincott, Williams &Wilkins, 2000; and Rowe, Raymond C. Handbook of PharmaceuticalExcipients. Chicago, Pharmaceutical Press, 2005. The formulations mayalso include one or more buffers, stabilizing agents, surfactants,wetting agents, lubricating agents, emulsifiers, suspending agents,preservatives, antioxidants, opaquing agents, glidants, processing aids,colorants, sweeteners, perfuming agents, flavoring agents, diluents andother known additives to provide an elegant presentation of the drug(i.e., a compound of the present invention or pharmaceutical compositionthereof) or aid in the manufacturing of the pharmaceutical product(i.e., medicament).

Suitable carriers, diluents and excipients are well known to thoseskilled in the art and include materials such as carbohydrates, waxes,water soluble and/or swellable polymers, hydrophilic or hydrophobicmaterials, gelatin, oils, solvents, water and the like. The particularcarrier, diluent or excipient used will depend upon the means andpurpose for which a compound of the present invention is being applied.Solvents are generally selected based on solvents recognized by personsskilled in the art as safe (GRAS) to be administered to a mammal. Ingeneral, safe solvents are non-toxic aqueous solvents such as water andother non-toxic solvents that are soluble or miscible in water. Suitableaqueous solvents include water, ethanol, propylene glycol, polyethyleneglycols (e.g., PEG 400, PEG 300), etc. and mixtures thereof. Theformulations can also include one or more buffers, stabilizing agents,surfactants, wetting agents, lubricating agents, emulsifiers, suspendingagents, preservatives, antioxidants, opaquing agents, glidants,processing aids, colorants, sweeteners, perfuming agents, flavoringagents and other known additives to provide an elegant presentation ofthe drug (i.e., a compound of the present invention or pharmaceuticalcomposition thereof) or aid in the manufacturing of the pharmaceuticalproduct (i.e., medicament).

Acceptable diluents, carriers, excipients and stabilizers are nontoxicto recipients at the dosages and concentrations employed, and includebuffers such as phosphate, citrate and other organic acids; antioxidantsincluding ascorbic acid and methionine; preservatives (such asoctadecyldimethylbenzyl ammonium chloride; hexamethonium chloride;benzalkonium chloride, benzethonium chloride; phenol, butyl or benzylalcohol; alkyl parabens such as methyl or propyl paraben; catechol;resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecularweight (less than about 10 residues) polypeptides; proteins, such asserum albumin, gelatin, or immunoglobulins; hydrophilic polymers such aspolyvinylpyrrolidone; amino acids such as glycine, glutamine,asparagine, histidine, arginine, or lysine; monosaccharides,disaccharides and other carbohydrates including glucose, mannose, ordextrins; chelating agents such as EDTA; sugars such as sucrose,mannitol, trehalose or sorbitol; salt-forming counter-ions such assodium; metal complexes (e.g., Zn-protein complexes); and/or non-ionicsurfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG). Aactive pharmaceutical ingredient of the invention (e.g., compound of fof the invention) can also be entrapped in microcapsules prepared, forexample, by coacervation techniques or by interfacial polymerization,for example, hydroxymethylcellulose or gelatin-microcapsules andpoly-(methylmethacylate) microcapsules, respectively, in colloidal drugdelivery systems (for example, liposomes, albumin microspheres,microemulsions, nano-particles and nanocapsules) or in macroemulsions.Such techniques are disclosed in Remington: The Science and Practice ofPharmacy: Remington the Science and Practice of Pharmacy (2005) 21^(St)Edition, Lippincott Williams & Wilkins, Philidelphia, Pa.

Sustained-release preparations of a compound can be prepared. Suitableexamples of sustained-release preparations include semipermeablematrices of solid hydrophobic polymers containing a compound asdescribed herein, which matrices are in the form of shaped articles,e.g., films, or microcapsules. Examples of sustained-release matricesinclude polyesters, hydrogels (for example,poly(2-hydroxyethyl-methacrylate), or poly(vinyl alcohol)), polylactides(U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid andgamma-ethyl-L-glutamate (Sidman et al., Biopolymers 22:547, 1983),non-degradable ethylene-vinyl acetate (Langer et al., J. Biomed. Mater.Res. 15:167, 1981), degradable lactic acid-glycolic acid copolymers suchas the LUPRON DEPOT™ (injectable microspheres composed of lacticacid-glycolic acid copolymer and leuprolide acetate) andpoly-D-(−)-3-hydroxybutyric acid (EP 133,988A). Sustained releasecompositions also include liposomally entrapped compounds, which can beprepared by methods known per se (Epstein et al., Proc. Natl. Acad. Sci.U.S.A. 82:3688, 1985; Hwang et al., Proc. Natl. Acad. Sci. U.S.A.77:4030, 1980; U.S. Pat. Nos. 4,485,045 and 4,544,545; and EP 102,324A).Ordinarily, the liposomes are of the small (about 200-800 Angstroms)unilamelar type in which the lipid content is greater than about 30 mol% cholesterol, the selected proportion being adjusted for the optimaltherapy.

The formulations include those suitable for the administration routesdetailed herein. The formulations can conveniently be presented in unitdosage form and can be prepared by any of the methods well known in theart of pharmacy. Techniques and formulations generally are found inRemington: The Science and Practice of Pharmacy: Remington the Scienceand Practice of Pharmacy (2005) 21^(st) Edition, Lippincott Williams &Wilkins, Philidelphia, Pa. Such methods include the step of bringinginto association the active ingredient with the carrier whichconstitutes one or more accessory ingredients.

In general the formulations are prepared by uniformly and intimatelybringing into association the active ingredient with liquid carriers,diluents or excipients or finely divided solid carriers, diluents orexcipients, or both, and then, if necessary, shaping the product. Atypical formulation is prepared by mixing a compound of the presentinvention and a carrier, diluent or excipient. The formulations can beprepared using conventional dissolution and mixing procedures. Forexample, the bulk drug substance (i.e., compound of the presentinvention or stabilized form of the compound (e.g., complex with acyclodextrin derivative or other known complexation agent) is dissolvedin a suitable solvent in the presence of one or more of the excipientsdescribed above. A compound of the present invention is typicallyformulated into pharmaceutical dosage forms to provide an easilycontrollable dosage of the drug and to enable patient compliance withthe prescribed regimen.

In one example, compounds as described herein may be formulated bymixing at ambient temperature at the appropriate pH, and at the desireddegree of purity, with physiologically acceptable carriers, i.e.,carriers that are non-toxic to recipients at the dosages andconcentrations employed into a galenical administration form. The pH ofthe formulation depends mainly on the particular use and theconcentration of compound, but preferably ranges anywhere from about 3to about 8. In one example, a compound of the invention is formulated inan acetate buffer, at pH 5. In another embodiment, a compound of theinvention is sterile. The compound may be stored, for example, as asolid or amorphous composition, as a lyophilized formulation or as anaqueous solution.

Formulations of a compound as described herein suitable for oraladministration can be prepared as discrete units such as pills,capsules, cachets or tablets each containing a predetermined amount of acompound of the invention.

Compressed tablets can be prepared by compressing in a suitable machinethe active ingredient in a free-flowing form such as a powder orgranules, optionally mixed with a binder, lubricant, inert diluent,preservative, surface active or dispersing agent. Molded tablets can bemade by molding in a suitable machine a mixture of the powdered activeingredient moistened with an inert liquid diluent. The tablets canoptionally be coated or scored and optionally are formulated so as toprovide slow or controlled release of the active ingredient therefrom.

Tablets, troches, lozenges, aqueous or oil suspensions, dispersiblepowders or granules, emulsions, hard or soft capsules, e.g., gelatincapsules, syrups or elixirs can be prepared for oral use. Formulationsof a compound o as described herein intended for oral use can beprepared according to any method known to the art for the manufacture ofpharmaceutical compositions and such compositions can contain one ormore agents including sweetening agents, flavoring agents, coloringagents and preserving agents, in order to provide a palatablepreparation. Tablets containing the active ingredient in admixture withnon-toxic pharmaceutically acceptable excipient which are suitable formanufacture of tablets are acceptable. These excipients can be, forexample, inert diluents, such as calcium or sodium carbonate, lactose,calcium or sodium phosphate; granulating and disintegrating agents, suchas maize starch, or alginic acid; binding agents, such as starch,gelatin or acacia; and lubricating agents, such as magnesium stearate,stearic acid or talc. Tablets can be uncoated or can be coated by knowntechniques including microencapsulation to delay disintegration andadsorption in the gastrointestinal tract and thereby provide a sustainedaction over a longer period. For example, a time delay material such asglyceryl monostearate or glyceryl distearate alone or with a wax can beemployed.

An example of a suitable oral administration form is a tablet containingabout 1 mg, 5 mg, 10 mg, 25 mg, 30 mg, 50 mg, 80 mg, 100 mg, 150 mg, 250mg, 300 mg and 500 mg of the compound of the invention compounded withabout 90-30 mg anhydrous lactose, about 5-40 mg sodium croscarmellose,about 5-30 mg polyvinylpyrrolidone (PVP) K30, and about 1-10 mgmagnesium stearate. The powdered ingredients are first mixed togetherand then mixed with a solution of the PVP. The resulting composition canbe dried, granulated, mixed with the magnesium stearate and compressedto tablet form using conventional equipment. An example of an aerosolformulation can be prepared by dissolving the compound, for example5-400 mg, of the invention in a suitable buffer solution, e.g. aphosphate buffer, adding a tonicifier, e.g. a salt such sodium chloride,if desired. The solution may be filtered, e.g., using a 0.2 micronfilter, to remove impurities and contaminants.

For treatment of the eye or other external tissues, e.g., mouth andskin, the formulations are preferably applied as a topical ointment orcream containing the active ingredient(s) in an amount of, for example,0.075 to 20% w/w. When formulated in an ointment, the active ingredientcan be employed with either a paraffinic or a water-miscible ointmentbase. Alternatively, the active ingredients can be formulated in a creamwith an oil-in-water cream base. If desired, the aqueous phase of thecream base can include a polyhydric alcohol, i.e., an alcohol having twoor more hydroxyl groups such as propylene glycol, butane 1,3-diol,mannitol, sorbitol, glycerol and polyethylene glycol (including PEG 400)and mixtures thereof. The topical formulations can desirably include acompound which enhances absorption or penetration of the activeingredient through the skin or other affected areas. Examples of suchdermal penetration enhancers include dimethyl sulfoxide and relatedanalogs.

The oily phase of the emulsions of this invention can be constitutedfrom known ingredients in a known manner. While the phase can comprisemerely an emulsifier, it desirably comprises a mixture of at least oneemulsifier with a fat or an oil or with both a fat and an oil.Preferably, a hydrophilic emulsifier is included together with alipophilic emulsifier which acts as a stabilizer. It is also preferredto include both an oil and a fat. Together, the emulsifier(s) with orwithout stabilizer(s) make up the so-called emulsifying wax, and the waxtogether with the oil and fat make up the so-called emulsifying ointmentbase which forms the oily dispersed phase of the cream formulations.Emulsifiers and emulsion stabilizers suitable for use in the formulationof the invention include Tween® 60, Span® 80, cetostearyl alcohol,benzyl alcohol, myristyl alcohol, glyceryl mono-stearate and sodiumlauryl sulfate.

In one aspect of topical applications, it is desired to administer aneffective amount of a pharmaceutical composition according to theinvention to target area, e.g., skin surfaces, mucous membranes, and thelike, which are adjacent to peripheral neurons which are to be treated.This amount will generally range from about 0.0001 mg to about 1 g of acompound of the invention per application, depending upon the area to betreated, whether the use is diagnostic, prophylactic or therapeutic, theseverity of the symptoms, and the nature of the topical vehicleemployed. A preferred topical preparation is an ointment, wherein about0.001 to about 50 mg of active ingredient is used per cc of ointmentbase. The pharmaceutical composition can be formulated as transdermalcompositions or transdermal delivery devices (“patches”). Suchcompositions include, for example, a backing, active compound reservoir,a control membrane, liner and contact adhesive. Such transdermal patchesmay be used to provide continuous pulsatile, or on demand delivery ofthe compounds of the present invention as desired.

Aqueous suspensions of a compound o as described herein contain theactive materials in admixture with excipients suitable for themanufacture of aqueous suspensions. Such excipients include a suspendingagent, such as sodium carboxymethylcellulose, croscarmellose, povidone,methylcellulose, hydroxypropyl methylcellulose, sodium alginate,polyvinylpyrrolidone, gum tragacanth and gum acacia, and dispersing orwetting agents such as a naturally occurring phosphatide (e.g.,lecithin), a condensation product of an alkylene oxide with a fatty acid(e.g., polyoxyethylene stearate), a condensation product of ethyleneoxide with a long chain aliphatic alcohol (e.g.,heptadecaethyleneoxycetanol), a condensation product of ethylene oxidewith a partial ester derived from a fatty acid and a hexitol anhydride(e.g., polyoxyethylene sorbitan monooleate). The aqueous suspension canalso contain one or more preservatives such as ethyl or n-propylp-hydroxybenzoate, one or more coloring agents, one or more flavoringagents and one or more sweetening agents, such as sucrose or saccharin.

Formulations of a compound as described herein can be in the form of asterile injectable preparation, such as a sterile injectable aqueous oroleaginous suspension. This suspension can be formulated according tothe known art using those suitable dispersing or wetting agents andsuspending agents which have been mentioned above. The sterileinjectable preparation can also be a sterile injectable solution orsuspension in a non-toxic parenterally acceptable diluent or solvent,such as a solution in 1,3-butanediol or prepared as a lyophilizedpowder. Among the acceptable vehicles and solvents that can be employedare water, Ringer's solution and isotonic sodium chloride solution. Inaddition, sterile fixed oils can conventionally be employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid can likewise be used in the preparation ofinjectables.

The amount of active ingredient that can be combined with the carriermaterial to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. For example, atime-release formulation intended for oral administration to humans cancontain approximately 1 to 1000 mg of active material compounded with anappropriate and convenient amount of carrier material which can varyfrom about 5 to about 95% of the total compositions (weight:weight). Thepharmaceutical composition can be prepared to provide easily measurableamounts for administration. For example, an aqueous solution intendedfor intravenous infusion can contain from about 3 to 500 μg of theactive ingredient per milliliter of solution in order that infusion of asuitable volume at a rate of about 30 mL/hr can occur.

Formulations suitable for parenteral administration include aqueous andnon-aqueous sterile injection solutions which can contain anti-oxidants,buffers, bacteriostats and solutes which render the formulation isotonicwith the blood of the intended recipient; and aqueous and non-aqueoussterile suspensions which can include suspending agents and thickeningagents.

Formulations suitable for topical administration to the eye also includeeye drops wherein the active ingredient is dissolved or suspended in asuitable carrier, especially an aqueous solvent for the activeingredient. The active ingredient is preferably present in suchformulations in a concentration of about 0.5 to 20% w/w, for exampleabout 0.5 to 10% w/w, for example about 1.5% w/w.

Formulations suitable for topical administration in the mouth includelozenges comprising the active ingredient in a flavored basis, usuallysucrose and acacia or tragacanth; pastilles comprising the activeingredient in an inert basis such as gelatin and glycerin, or sucroseand acacia; and mouthwashes comprising the active ingredient in asuitable liquid carrier.

Formulations for rectal administration can be presented as a suppositorywith a suitable base comprising for example cocoa butter or asalicylate.

Formulations suitable for intrapulmonary or nasal administration have aparticle size for example in the range of 0.1 to 500 microns (includingparticle sizes in a range between 0.1 and 500 microns in incrementsmicrons such as 0.5, 1, 30 microns, 35 microns, etc.), which isadministered by rapid inhalation through the nasal passage or byinhalation through the mouth so as to reach the alveolar sacs. Suitableformulations include aqueous or oily solutions of the active ingredient.Formulations suitable for aerosol or dry powder administration can beprepared according to conventional methods and can be delivered withother therapeutic agents such as compounds heretofore used in thetreatment of disorders as described below.

The formulations can be packaged in unit-dose or multi-dose containers,for example sealed ampoules and vials, and can be stored in afreeze-dried (lyophilized) condition requiring only the addition of thesterile liquid carrier, for example water, for injection immediatelyprior to use. Extemporaneous injection solutions and suspensions areprepared from sterile powders, granules and tablets of the kindpreviously described. Preferred unit dosage formulations are thosecontaining a daily dose or unit daily sub-dose, as herein above recited,or an appropriate fraction thereof, of the active ingredient.

When the binding target is located in the brain, certain embodiments ofthe invention provide for a compound as described herein to traverse theblood-brain barrier. Certain neurodegenerative diseases are associatedwith an increase in permeability of the blood-brain barrier, such that acompound of of the invention can be readily introduced to the brain.When the blood-brain barrier remains intact, several art-knownapproaches exist for transporting molecules across it, including, butnot limited to, physical methods, lipid-based methods, and receptor andchannel-based methods.

Physical methods of transporting a compound o as described herein acrossthe blood-brain barrier include, but are not limited to, circumventingthe blood-brain barrier entirely, or by creating openings in theblood-brain barrier.

Circumvention methods include, but are not limited to, direct injectioninto the brain (see, e.g., Papanastassiou et al., Gene Therapy9:398-406, 2002), interstitial infusion/convection-enhanced delivery(see, e.g., Bobo et al., Proc. Natl. Acad. Sci. U.S.A. 91:2076-2080,1994), and implanting a delivery device in the brain (see, e.g., Gill etal., Nature Med. 9:589-595, 2003; and Gliadel Wafers™, GuildfordPharmaceutical). Methods of creating openings in the barrier include,but are not limited to, ultrasound (see, e.g., U.S. Patent PublicationNo. 2002/0038086), osmotic pressure (e.g., by administration ofhypertonic mannitol (Neuwelt, E. A., Implication of the Blood-BrainBarrier and its Manipulation, Volumes 1 and 2, Plenum Press, N.Y.,1989)), and permeabilization by, e.g., bradykinin or permeabilizer A-7(see, e.g., U.S. Pat. Nos. 5,112,596, 5,268,164, 5,506,206, and5,686,416).

Lipid-based methods of transporting a compound o as described hereinacross the blood-brain barrier include, but are not limited to,encapsulating the a compound o as described herein in liposomes that arecoupled to antibody binding fragments that bind to receptors on thevascular endothelium of the blood-brain barrier (see, e.g., U.S. PatentApplication Publication No. 2002/0025313), and coating a compound asdescribed herein in low-density lipoprotein particles (see, e.g., U.S.Patent Application Publication No. 2004/0204354) or apolipoprotein E(see, e.g., U.S. Patent Application Publication No. 2004/0131692).

Receptor and channel-based methods of transporting a compound o asdescribed herein across the blood-brain barrier include, but are notlimited to, using glucocorticoid blockers to increase permeability ofthe blood-brain barrier (see, e.g., U.S. Patent Application PublicationNos. 2002/0065259, 2003/0162695, and 2005/0124533); activating potassiumchannels (see, e.g., U.S. Patent Application Publication No.2005/0089473), inhibiting ABC drug transporters (see, e.g., U.S. PatentApplication Publication No. 2003/0073713); coating a compound o asdescribed herein with a transferrin and modulating activity of the oneor more transferrin receptors (see, e.g., U.S. Patent ApplicationPublication No. 2003/0129186), and cationizing the antibodies (see,e.g., U.S. Pat. No. 5,004,697).

For intracerebral use, in certain embodiments, the compounds can beadministered continuously by infusion into the fluid reservoirs of theCNS, although bolus injection may be acceptable. The inhibitors can beadministered into the ventricles of the brain or otherwise introducedinto the CNS or spinal fluid. Administration can be performed by use ofan indwelling catheter and a continuous administration means such as apump, or it can be administered by implantation, e.g., intracerebralimplantation of a sustained-release vehicle. More specifically, theinhibitors can be injected through chronically implanted cannulas orchronically infused with the help of osmotic minipumps. Subcutaneouspumps are available that deliver proteins through a small tubing to thecerebral ventricles. Highly sophisticated pumps can be refilled throughthe skin and their delivery rate can be set without surgicalintervention. Examples of suitable administration protocols and deliverysystems involving a subcutaneous pump device or continuousintracerebroventricular infusion through a totally implanted drugdelivery system are those used for the administration of dopamine,dopamine agonists, and cholinergic agonists to Alzheimer's diseasepatients and animal models for Parkinson's disease, as described byHarbaugh, J. Neural Transm. Suppl. 24:271, 1987; and DeYebenes et al.,Mov. Disord. 2: 143, 1987.

A compound as described herein used in the invention are formulated,dosed, and administered in a fashion consistent with good medicalpractice. Factors for consideration in this context include theparticular disorder being treated, the particular mammal being treated,the clinical condition of the individual patient, the cause of thedisorder, the site of delivery of the agent, the method ofadministration, the scheduling of administration, and other factorsknown to medical practitioners. A compound as described herein need notbe, but is optionally formulated with one or more agent currently usedto prevent or treat the disorder in question. The effective amount ofsuch other agents depends on the amount of a compound of the inventionpresent in the formulation, the type of disorder or treatment, and otherfactors discussed above.

These are generally used in the same dosages and with administrationroutes as described herein, or about from 1 to 99% of the dosagesdescribed herein, or in any dosage and by any route that isempirically/clinically determined to be appropriate.

For the prevention or treatment of disease, the appropriate dosage of acompound as described herein (when used alone or in combination withother agents) will depend on the type of disease to be treated, theproperties of the compound, the severity and course of the disease,whether the compound is administered for preventive or therapeuticpurposes, previous therapy, the patient's clinical history and responseto the compound, and the discretion of the attending physician. Thecompound is suitably administered to the patient at one time or over aseries of treatments. Depending on the type and severity of the disease,about 1 μg/kg to 15 mg/kg (e.g., 0.1 mg/kg-10 mg/kg) of compound can bean initial candidate dosage for administration to the patient, whether,for example, by one or more separate administrations, or by continuousinfusion. One typical daily dosage might range from about 1 μg kg to 100mg/kg or more, depending on the factors mentioned above. For repeatedadministrations over several days or longer, depending on the condition,the treatment would generally be sustained until a desired suppressionof disease symptoms occurs. One exemplary dosage of a compound of theinvention would be in the range from about 0.05 mg/kg to about 10 mg/kg.Thus, one or more doses of about 0.5 mg/kg, 2.0 mg/kg, 4.0 mg/kg, or 10mg/kg (or any combination thereof) may be administered to the patient.Such doses may be administered intermittently, e.g., every week or everythree weeks (e.g., such that the patient receives from about two toabout twenty, or, e.g., about six doses of the antibody). An initialhigher loading dose, followed by one or more lower doses may beadministered. An exemplary dosing regimen comprises administering aninitial loading dose of about 4 mg/kg, followed by a weekly maintenancedose of about 2 mg kg of the compound. However, other dosage regimensmay be useful. The progress of this therapy is easily monitored byconventional techniques and assays.

Other typical daily dosages might range from, for example, about 1 g/kgto up to 100 mg/kg or more (e.g., about 1 μg kg to 1 mg/kg, about 1μg/kg to about 5 mg/kg, about 1 mg kg to 10 mg/kg, about 5 mg/kg toabout 200 mg/kg, about 50 mg/kg to about 150 mg/mg, about 100 mg/kg toabout 500 mg/kg, about 100 mg/kg to about 400 mg/kg, and about 200 mg/kgto about 400 mg/kg), depending on the factors mentioned above.Typically, the clinician will administer a compound until a dosage isreached that results in improvement in or, optimally, elimination of,one or more symptoms of the treated disease or condition. The progressof this therapy is easily monitored by conventional assays. One or moreagent provided herein may be administered together or at different times(e.g., one agent is administered prior to the administration of a secondagent). One or more agent may be administered to a subject usingdifferent techniques (e.g., one agent may be administered orally, whilea second agent is administered via intramuscular injection orintranasally). One or more agent may be administered such that the oneor more agent has a pharmacologic effect in a subject at the same time.Alternatively, one or more agent may be administered, such that thepharmacological activity of the first administered agent is expiredprior the administration of one or more secondarily administered agents(e.g., 1, 2, 3, or 4 secondarily administered agents).

Indications and Methods of Treatment

The compounds of the invention modulate, preferably inhibit, ion fluxthrough a voltage-dependent sodium channel in a mammal, (e.g, a human).Any such modulation, whether it be partial or complete inhibition orprevention of ion flux, is sometimes referred to herein as “blocking”and corresponding compounds as “blockers” or “inhibitors”. In general,the compounds of the invention modulate the activity of a sodium channeldownwards by inhibiting the voltage-dependent activity of the sodiumchannel, and/or reduce or prevent sodium ion flux across a cell membraneby preventing sodium channel activity such as ion flux.

Accordingly, the compounds of the invention are sodium channel blockersand are therefore useful for treating diseases and conditions inmammals, for example humans, and other organisms, including all thosediseases and conditions which are the result of aberrantvoltage-dependent sodium channel biological activity or which may beameliorated by modulation of voltage-dependent sodium channel biologicalactivity. In particular, the compounds of the invention, i.e., thecompounds of formula (I) and embodiments and (or stereoisomers,geometric isomers, tautomers, solvates, metabolites, isotopes,pharmaceutically acceptable salts, or prodrugs thereof), are useful fortreating diseases and conditions in mammals, for example humans, whichare the result of aberrant voltage-dependent NaV1.7 biological activityor which may be ameliorated by the modulation, preferably theinhibition, of NaV1.7 biological activity. In certain aspects, thecompounds of the invention selectively inhibit NaV1.7 over NaV1.5.

As defined herein, a sodium channel-mediated disease or condition refersto a disease or condition in a mammal, preferably a human, which isameliorated upon modulation of the sodium channel and includes, but isnot limited to, pain, central nervous conditions such as epilepsy,anxiety, depression and bipolar disease; cardiovascular conditions suchas arrhythmias, atrial fibrillation and ventricular fibrillation;neuromuscular conditions such as restless leg syndrome and muscleparalysis or tetanus; neuroprotection against stroke, neural trauma andmultiple sclerosis; and channelopathies such as erythromyalgia andfamilial rectal pain syndrome.

In one aspect, the present invention relates to compounds,pharmaceutical compositions and methods of using the compounds andpharmaceutical compositions for the treatment of sodium channel-mediateddiseases in mammals, preferably humans and preferably diseases andconditions related to pain, central nervous conditions such as epilepsy,anxiety, depression and bipolar disease; cardiovascular conditions suchas arrhythmias, atrial fibrillation and ventricular fibrillation;neuromuscular conditions such as restless leg syndrome and muscleparalysis or tetanus; neuroprotection against stroke, neural trauma andmultiple sclerosis; and channelopathies such as erythromyalgia andfamilial rectal pain syndrome, by administering to a mammal, for examplea human, in need of such treatment an effective amount of a sodiumchannel blocker modulating, especially inhibiting, agent.

A sodium channel-mediated disease or condition also includes painassociated with HIV, HIV treatment induced neuropathy, trigeminalneuralgia, glossopharyngeal neuralgia, neuropathy secondary tometastatic infiltration, adiposis dolorosa, thalamic lesions,hypertension, autoimmune disease, asthma, drug addiction (e.g., opiate,benzodiazepine, amphetamine, cocaine, alcohol, butane inhalation),Alzheimer, dementia, age-related memory impairment, Korsakoff syndrome,restenosis, urinary dysfunction, incontinence, Parkinson's disease,cerebrovascular ischemia, neurosis, gastrointestinal disease, sicklecell anemia, transplant rejection, heart failure, myocardial infarction,reperfusion injury, intermittant claudication, angina, convulsion,respiratory disorders, cerebral or myocardial ischemias, long-QTsyndrome, Catecholeminergic polymorphic ventricular tachycardia,ophthalmic diseases, spasticity, spastic paraplegia, myopathies,myasthenia gravis, paramyotonia congentia, hyperkalemic periodicparalysis, hypokalemic periodic paralysis, alopecia, anxiety disorders,psychotic disorders, mania, paranoia, seasonal affective disorder, panicdisorder, obsessive compulsive disorder (OCD), phobias, autism,Aspergers Syndrome, Retts syndrome, disintegrative disorder, attentiondeficit disorder, aggressivity, impulse control disorders, thrombosis,pre clampsia, congestive cardiac failure, cardiac arrest, Freidrich'sataxia, Spinocerebellear ataxia, myelopathy, radiculopathy, systemiclupus erythamatosis, granulomatous disease, olivo-ponto-cerebellaratrophy, spinocerebellar ataxia, episodic ataxia, myokymia, progressivepallidal atrophy, progressive supranuclear palsy and spasticity,traumatic brain injury, cerebral oedema, hydrocephalus injury, spinalcord injury, anorexia nervosa, bulimia, Prader-Willi syndrome, obesity,optic neuritis, cataract, retinal haemorrhage, ischaemic retinopathy,retinitis pigmentosa, acute and chronic glaucoma, macular degeneration,retinal artery occlusion, Chorea, Huntington's chorea, cerebral edema,proctitis, post-herpetic neuralgia, eudynia, heat sensitivity,sarcoidosis, irritable bowel syndrome, Tourette syndrome, Lesch-NyhanSyndrome, Brugado syndrome, Liddle syndrome, Crohns disease, multiplesclerosis and the pain associated with multiple sclerosis (MS),amyotrophic lateral sclerosis (ALS), disseminated sclerosis, diabeticneuropathy, peripheral neuropathy, charcot marie tooth syndrome,arthritic, rheumatoid arthritis, osteoarthritis, chondrocalcinosis,atherosclerosis, paroxysmal dystonia, myasthenia syndromes, myotonia,myotonic dystrophy, muscular dystrophy, malignant hyperthermia, cysticfibrosis, pseudoaldosteronism, rhabdomyolysis, mental handicap,hypothyroidism, bipolar depression, anxiety, schizophrenia, sodiumchannel toxin related illnesses, familial erythromelalgia, primaryerythromelalgia, rectal pain, cancer, epilepsy, partial and generaltonic seizures, febrile seizures, absence seizures (petit mal),myoclonic seizures, atonic seizures, clonic seizures, Lennox Gastaut,West Syndome (infantile spasms), multiresistant seizures, seizureprophylaxis (anti-epileptogenic), familial Mediterranean fever syndrome,gout, restless leg syndrome, arrhythmias, fibromyalgia, neuroprotectionunder ischaemic conditions caused by stroke or neural trauma,tachy-arrhythmias, atrial fibrillation and ventricular fibrillation andas a general or local anaesthetic.

As used herein, the term “pain” refers to all categories of pain and isrecognized to include, but is not limited to, neuropathic pain,inflammatory pain, nociceptive pain, idiopathic pain, neuralgic pain,orofacial pain, burn pain, burning mouth syndrome, somatic pain,visceral pain, myofacial pain, dental pain, cancer pain, chemotherapypain, trauma pain, surgical pain, post-surgical pain, childbirth pain,labor pain, chronic regional pain syndrome (CRPS), reflex sympatheticdystrophy, brachial plexus avulsion, neurogenic bladder, acute pain(e.g., musculoskeletal and post-operative pain), chronic pain,persistent pain, peripherally mediated pain, centrally mediated pain,chronic headache, migraine headache, familial hemiplegic migraine,conditions associated with cephalic pain, sinus headache, tensionheadache, phantom limb pain, peripheral nerve injury, pain followingstroke, thalamic lesions, radiculopathy, HIV pain, post-herpetic pain,non-cardiac chest pain, irritable bowel syndrome and pain associatedwith bowel disorders and dyspepsia, and combinations thereof.

Furthermore, sodium channel blockers have clinical uses in addition topain. The present invention therefore also relates to compounds,pharmaceutical compositions and methods of using the compounds andpharmaceutical compositions for the treatment of diseases or conditionssuch as cancer and pruritus (itch).

Pruritus, commonly known as itch, is a common dermatological condition.While the exact causes of pruritus are complex and incompletelyunderstood, there has long been evidence that itch involves sensoryneurons, especially C fibers, similar to those that mediate pain(Schmelz, M., et al., J. Neurosci. (1997), 17: 8003-8). In particular,it is believed that sodium influx through voltage-gated sodium channelsis essential for the propagation of itch sensation from the skin.Transmission of the itch impulses results in the unpleasant sensationthat elicits the desire or reflex to scratch.

Multiple causes and electrical pathways for eliciting itch are known. Inhumans, pruritus can be elicited by histamine or PAR-2 agonists such asmucunain that activate distinct populations of C fibers (Namer, B., etal., J. Neurophysiol. (2008), 100: 2062-9). A variety of neurotrophicpeptides are known to mediate itch in animal models (Wang, H., andYosipovitch, G., International Journal of Dermatology (2010), 49: 1-11).Itch can also be elicited by opioids, evidence of distinct pharmacologyfrom that of pain responses.

There exists a complex interaction between itch and pain responses thatarises in part from the overlapping sensory input from the skin (Ikoma,A., et al., Arch. Dermatol. (2003), 139: 1475-8) and also from thediverse etiology of both pain and pruritus. Pain responses canexacerbate itching by enhancing central sensitization or lead toinhibition of painful scratching. Particularly severe forms of chronicitch occur when pain responses are absent, as in the case ofpost-herpetic itch (Oaklander, A. L., et al., Pain (2002), 96: 9-12).

The compounds of the invention can also be useful for treating pruritus.The rationale for treating itch with inhibitors of voltage-gated sodiumchannels, especially NaV1.7, is as follows: The propagation ofelectrical activity in the C fibers that sense pruritinergic stimulantsrequires sodium entry through voltage-gated sodium channels.

NaV1.7 is expressed in the C fibers and kerotinocytes in human skin(Zhao, P., et al., Pain (2008), 139: 90-105).

A gain of function mutation of NaV1.7 (L858F) that causeserythromelalgia also causes chronic itch (Li, Y., et al., Clinical andExperimental Dermatology (2009), 34: e313-e4).

Chronic itch can be alleviated with treatment by sodium channelblockers, such as the local anesthetic lidocaine (Oaklander, A. L., etal., Pain (2002), 96: 9-12; Villamil, A. G., et al., The AmericanJournal of Medicine (2005), 118: 1160-3). In these reports, lidocainewas effective when administered either intravenously or topically (aLidoderm patch). Lidocaine can have multiple activities at the plasmaconcentrations achieved when administered systemically, but whenadministered topically, the plasma concentrations are only about 1 μM(Center for Drug Evaluation and Research NDA 20-612). At theseconcentrations, lidocaine is selective for sodium channel block andinhibits spontaneous electrical activity in C fibers and pain responsesin animal models (Xiao, W. H., and Bennett, G. J. Pain (2008), 137:218-28). The types of itch or skin irritation, include, but are notlimited to:

psoriatic pruritus, itch due to hemodyalisis, aguagenic pruritus, anditching caused by skin disorders (e.g., contact dermatitis), systemicdisorders, neuropathy, psychogenic factors or a mixture thereof;

itch caused by allergic reactions, insect bites, hypersensitivity (e.g.,dry skin, acne, eczema, psoriasis), inflammatory conditions or injury;

itch associated with vulvar vestibulitis; and

skin irritation or inflammatory effect from administration of anothertherapeutic such as, for example, antibiotics, antivirals andantihistamines.

The compounds of the invention are also useful in treating certaincancers, such as hormone sensitive cancers, such as prostate cancer(adenocarcinoma), breast cancer, ovarian cancer, testicular cancer andthyroid neoplasia, in a mammal, preferably a human. The voltage gatedsodium channels have been demonstrated to be expressed in prostate andbreast cancer cells. Up-regulation of neonatal NaV1.5 occurs as anintegral part of the metastatic process in human breast cancer and couldserve both as a novel marker of the metastatic phenotype and atherapeutic target (Clin. Cancer Res. (2005), August 1; 11(15): 5381-9).Functional expression of voltage-gated sodium channel alpha-subunits,specifically NaV1.7, is associated with strong metastatic potential inprostate cancer (CaP) in vitro. Voltage-gated sodium channelalpha-subunits immunostaining, using antibodies specific to the sodiumchannel alpha subunit was evident in prostatic tissues and markedlystronger in CaP vs non-CaP patients (Prostate Cancer Prostatic Dis.,2005; 8(3):266-73). See also Diss, J. K. J., et al., Mol. Cell.Neurosci. (2008), 37:537-547 and Kis-Toth, K., et al., The Journal ofImmunology (2011), 187:1273-1280.

In consideration of the above, in one embodiment, the present inventionprovides a method for treating a mammal for, or protecting a mammal fromdeveloping, a sodium channel-mediated disease, especially pain,comprising administering to the mammal, especially a human, in needthereof, a therapeutically effective amount of a compound of theinvention or a pharmaceutical composition comprising a therapeuticallyeffective amount of a compound of the invention wherein the compoundmodulates the activity of one or more voltage-dependent sodium channels.

In another embodiment of the invention is a method of treating a diseaseor a condition in a mammal, preferably a human, wherein the disease orcondition is selected from the group consisting of pain, depression,cardiovascular diseases, respiratory diseases, and psychiatric diseases,and combinations thereof, and wherein the method comprises administeringto the mammal in need thereof a therapeutically effective amount of anembodiment of a compound of the invention, as set forth above, as astereoisomer, enantiomer or tautomer thereof or mixtures thereof, or apharmaceutically acceptable salt, solvate or prodrug thereof, or apharmaceutical composition comprising a therapeutically effective amountof a compound of the invention, as set forth above, as a stereoisomer,enantiomer or tautomer thereof or mixtures thereof, or apharmaceutically acceptable salt, solvate or prodrug thereof, and apharmaceutically acceptable excipient.

One embodiment of this embodiment is wherein the disease or condition isselected from the group consisting of acute pain, chronic pain,neuropathic pain, inflammatory pain, visceral pain, cancer pain,chemotherapy pain, trauma pain, surgical pain, post surgical pain,childbirth pain, labor pain, neurogenic bladder, ulcerative colitis,persistent pain, peripherally mediated pain, centrally mediated pain,chronic headache, migraine headache, sinus headache, tension headache,phantom limb pain, peripheral nerve injury, and combinations thereof.

Another embodiment of this embodiment is wherein the disease orcondition is selected from the group consisting of pain associated withHIV, HIV treatment induced neuropathy, trigeminal neuralgia, postherpetic neuralgia, eudynia, heat sensitivity, tosarcoidosis, irritablebowel syndrome, Crohns disease, pain associated with multiple sclerosis(MS), amyotrophic lateral sclerosis (ALS), diabetic neuropathy,peripheral neuropathy, arthritic, rheumatoid arthritis, osteoarthritis,atherosclerosis, paroxysmal dystonia, myasthenia syndromes, myotonia,malignant hyperthermia, cystic fibrosis, pseudoaldosteronism,rhabdomyolysis, hypothyroidism, bipolar depression, anxiety,schizophrenia, sodium channel toxin related illnesses, familialerythromelalgia, primary erythromelalgia, familial rectal pain, cancer,epilepsy, partial and general tonic seizures, restless leg syndrome,arrhythmias, fibromyalgia, neuroprotection under ischaemic conditionscaused by stroke or neural trauma, tachy arrhythmias, atrialfibrillation and ventricular fibrillation.

Another embodiment of the invention is a method of treating, but notpreventing, pain in a mammal, wherein the method comprises administeringto the mammal in need thereof a therapeutically effective amount of acompound of the invention, as set forth above, as a stereoisomer,enantiomer or tautomer thereof or mixtures thereof, or apharmaceutically acceptable salt, solvate or prodrug thereof, or apharmaceutical composition comprising a therapeutically effective amountof a compound of the invention, as set forth above, as a stereoisomer,enantiomer or tautomer thereof or mixtures thereof, or apharmaceutically acceptable salt, solvate or prodrug thereof, and apharmaceutically acceptable excipient.

One embodiment of this embodiment is a method wherein the pain isselected from the group consisting of neuropathic pain, inflammatorypain, visceral pain, cancer pain, chemotherapy pain, trauma pain,surgical pain, post surgical pain, childbirth pain, labor pain, dentalpain, chronic pain, persistent pain, peripherally mediated pain,centrally mediated pain, chronic headache, migraine headache, sinusheadache, tension headache, phantom limb pain, peripheral nerve injury,trigeminal neuralgia, post herpetic neuralgia, eudynia, familialerythromelalgia, primary erythromelalgia, familial rectal pain orfibromyalgia, and combinations thereof.

Another embodiment of this embodiment is a method wherein the pain isassociated with a disease or condition selected from HIV, HIV treatmentinduced neuropathy, heat sensitivity, tosarcoidosis, irritable bowelsyndrome, Crohns disease, multiple sclerosis, amyotrophic lateralsclerosis, diabetic neuropathy, peripheral neuropathy, rheumatoidarthritis, osteoarthritis, atherosclerosis, paroxysmal dystonia,myasthenia syndromes, myotonia, malignant hyperthermia, cystic fibrosis,pseudoaldosteronism, rhabdomyolysis, hypothyroidism, bipolar depression,anxiety, schizophrenia, sodium channel toxin related illnesses,neurogenic bladder, ulcerative colitis, cancer, epilepsy, partial andgeneral tonic seizures, restless leg syndrome, arrhythmias, ischaemicconditions caused by stroke or neural trauma, tachy arrhythmias, atrialfibrillation and ventricular fibrillation.

Another embodiment of the invention is the method of treating pain in amammal, preferably a human, by the inhibition of ion flux through avoltage dependent sodium channel in the mammal, wherein the methodcomprises administering to the mammal in need thereof a therapeuticallyeffective amount of an embodiment of a compound of the invention, as setforth above, as a stereoisomer, enantiomer or tautomer thereof ormixtures thereof, or a pharmaceutically acceptable salt, solvate orprodrug thereof, or a pharmaceutical composition comprising atherapeutically effective amount of a compound of the invention, as setforth above, as a stereoisomer, enantiomer or tautomer thereof ormixtures thereof, or a pharmaceutically acceptable salt, solvate orprodrug thereof, and a pharmaceutically acceptable excipient.

Another embodiment of the invention is the method of treating pruritusin a mammal, preferably a human, wherein the method comprisesadministering to the mammal in need thereof a therapeutically effectiveamount of an embodiment of a compound of the invention, as set forthabove, as a stereoisomer, enantiomer or tautomer thereof or mixturesthereof, or a pharmaceutically acceptable salt, solvate or prodrugthereof, or a pharmaceutical composition comprising a therapeuticallyeffective amount of a compound of the invention, as set forth above, asa stereoisomer, enantiomer or tautomer thereof or mixtures thereof, or apharmaceutically acceptable salt, solvate or prodrug thereof, and apharmaceutically acceptable excipient.

Another embodiment of the invention is the method of treating cancer ina mammal, preferably a human, wherein the method comprises administeringto the mammal in need thereof a therapeutically effective amount of anembodiment of a compound of the invention, as set forth above, as astereoisomer, enantiomer or tautomer thereof or mixtures thereof, or apharmaceutically acceptable salt, solvate or prodrug thereof, or apharmaceutical composition comprising a therapeutically effective amountof a compound of the invention, as set forth above, as a stereoisomer,enantiomer or tautomer thereof or mixtures thereof, or apharmaceutically acceptable salt, solvate or prodrug thereof, and apharmaceutically acceptable excipient.

Another embodiment of the invention is the method of decreasing ion fluxthrough a voltage dependent sodium channel in a cell in a mammal,wherein the method comprises contacting the cell with an embodiment of acompound of the invention, as set forth above, as a stereoisomer,enantiomer or tautomer thereof or mixtures thereof, or apharmaceutically acceptable salt, solvate or prodrug thereof.

Another embodiment of the invention is the method of selectivelyinhibiting a first voltage-gated sodium channel over a secondvoltage-gated sodium channel in a mammal, wherein the method comprisesadministering to the mammal an inhibitory amount of a compound offormula (I), or an embodiment of a compound of formula (I).

Another embodiment of the invention is the method of selectivelyinhibiting NaV1.7 in a mammal or a mammalian cell as compared to NaV1.5,wherein the method comprises administering to the mammal in need thereofan inhibitory amount of a compound of formula (I) or an embodiment of anembodiment thereof.

For each of the above embodiments described related to treating diseasesand conditions in a mammal, the present invention also contemplatesrelatedly a compound as described herein for the use as a medicament inthe treatment of such diseases and conditions.

For each of the above embodiments described related to treating diseasesand conditions in a mammal, the present invention also contemplatesrelatedly the use of a compound o as described herein for themanufacture of a medicament for the treatment of such diseases andconditions.

Another embodiment of the invention is a method of using a compound asdescribed herein as a standard or control in in vitro or in vivo assaysin determining the efficacy of test compounds in modulatingvoltage-dependent sodium channels.

In another embodiment of the invention, the compounds as describedherein are isotopically-labeled by having one or more atoms thereinreplaced by an atom having a different atomic mass or mass number. Suchisotopically-labeled (i.e., radiolabelled) compounds are considered tobe within the scope of this invention. Examples of isotopes that can beincorporated into the compounds include isotopes of hydrogen, carbon,nitrogen, oxygen, phosphorous, sulfur, fluorine, chlorine, and iodine,such as, but not limited to, ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹³N, ⁵N, ¹⁵O, ¹⁷O,¹⁸O, ³¹P, ³²P, ³⁵S, ¹⁸F, ³⁶Cl, ¹²³I, and ¹²⁵I, respectively. Theseisotopically-labeled compounds would be useful to help determine ormeasure the effectiveness of the compounds, by characterizing, forexample, the site or mode of action on the sodium channels, or bindingaffinity to pharmacologically important site of action on the sodiumchannels, particularly NaV1.7. Certain isotopically-labeled compounds,for example, those incorporating a radioactive isotope, are useful indrug and/or substrate tissue distribution studies. The radioactiveisotopes tritium, i.e. ³H, and carbon-14, i.e., ¹⁴C, are particularlyuseful for this purpose in view of their ease of incorporation and readymeans of detection.

Substitution with heavier isotopes such as deuterium, i.e. ²H, mayafford certain therapeutic advantages resulting from greater metabolicstability, for example, increased in vivo half-life or reduced dosagerequirements, and hence may be preferred in some circumstances.

Substitution with positron emitting isotopes, such as ¹¹C, ¹⁸F, ¹⁵O and¹³N, can be useful in Positron Emission Topography (PET) studies forexamining substrate receptor occupancy. Isotopically-labeled compoundscan generally be prepared by conventional techniques known to thoseskilled in the art or by processes analogous to those described in theExamples as set out below using an appropriate isotopically-labeledreagent in place of the non-labeled reagent previously employed.

Testing Compounds

The assessment of the compounds of the invention in mediating,especially inhibiting, the sodium channel ion flux can be determinedusing the assays described hereinbelow. Alternatively, the assessment ofthe compounds in treating conditions and diseases in humans may beestablished in industry standard animal models for demonstrating theefficacy of compounds in treating pain. Animal models of humanneuropathic pain conditions have been developed that result inreproducible sensory deficits (allodynia, hyperalgesia, and spontaneouspain) over a sustained period of time that can be evaluated by sensorytesting. By establishing the degree of mechanical, chemical, andtemperature induced allodynia and hyperalgesia present, severalphysiopathological conditions observed in humans can be modeled allowingthe evaluation of pharmacotherapies.

In rat models of peripheral nerve injury, ectopic activity in theinjured nerve corresponds to the behavioural signs of pain. In thesemodels, intravenous application of the sodium channel blocker and localanesthetic lidocaine can suppress the ectopic activity and reverse thetactile allodynia at concentrations that do not affect general behaviourand motor function (Mao, J. and Chen, L. L, Pain (2000), 87:7-17).Allometric scaling of the doses effective in these rat models,translates into doses similar to those shown to be efficacious in humans(Tanelian, D. L. and Brose, W. G., Anesthesiology (1991),74(5):949-951). Furthermore, Lidoderm®, lidocaine applied in the form ofa dermal patch, is currently an FDA approved treatment for post-herpeticneuralgia (Devers, A. and Glaler, B. S., Clin. J. Pain (2000),16(3):205-8).

The present invention readily affords many different means foridentification of sodium channel modulating agents that are useful astherapeutic agents. Identification of modulators of sodium channel canbe assessed using a variety of in vitro and in vivo assays, e.g.,measuring current, measuring membrane potential, measuring ion flux,(e.g., sodium or guanidinium), measuring sodium concentration, measuringsecond messengers and transcription levels, and using e.g.,voltage-sensitive dyes, radioactive tracers, and patch-clampelectrophysiology.

One such protocol involves the screening of chemical agents for abilityto modulate the activity of a sodium channel thereby identifying it as amodulating agent.

A typical assay described in Bean et al., J. General Physiology (1983),83:613-642, and Leuwer, M., et al., Br. J. Pharmacol (2004),141(1):47-54, uses patch-clamp techniques to study the behaviour ofchannels. Such techniques are known to those skilled in the art, and maybe developed, using current technologies, into low or medium throughputassays for evaluating compounds for their ability to modulate sodiumchannel behaviour.

Throughput of test compounds is an important consideration in the choiceof screening assay to be used. In some strategies, where hundreds ofthousands of compounds are to be tested, it is not desirable to use lowthroughput means. In other cases, however, low throughput issatisfactory to identify important differences between a limited numberof compounds. Often it will be necessary to combine assay types toidentify specific sodium channel modulating compounds.

Electrophysiological assays using patch clamp techniques is accepted asa gold standard for detailed characterization of sodium channel compoundinteractions, and as described in Bean et al., op. cit. and Leuwer, M.,et al., op. cit. There is a manual low-throughput screening (LTS) methodwhich can compare 2-10 compounds per day; a recently developed systemfor automated medium-throughput screening (MTS) at 20-50 patches (i.e.compounds) per day; and a technology from Molecular Devices Corporation(Sunnyvale, Calif.) which permits automated high-throughput screening(HTS) at 1000-3000 patches (i.e. compounds) per day.

One automated patch-clamp system utilizes planar electrode technology toaccelerate the rate of drug discovery. Planar electrodes are capable ofachieving high-resistance, cells-attached seals followed by stable,low-noise whole-cell recordings that are comparable to conventionalrecordings. A suitable instrument is the PatchXpress 7000A (AxonInstruments Inc, Union City, Calif.). A variety of cell lines andculture techniques, which include adherent cells as well as cellsgrowing spontaneously in suspension are ranked for seal success rate andstability. Immortalized cells (e.g. HEK and CHO) stably expressing highlevels of the relevant sodium ion channel can be adapted intohigh-density suspension cultures.

Other assays can be selected which allow the investigator to identifycompounds which block specific states of the channel, such as the openstate, closed state or the resting state, or which block transition fromopen to closed, closed to resting or resting to open. Those skilled inthe art are generally familiar with such assays.

Binding assays are also available. Designs include traditionalradioactive filter based binding assays or the confocal basedfluorescent system available from Evotec OAI group of companies(Hamburg, Germany), both of which are HTS.

Radioactive flux assays can also be used. In this assay, channels arestimulated to open with veratridine or aconitine and held in astabilized open state with a toxin, and channel blockers are identifiedby their ability to prevent ion influx. The assay can use radioactive22[Na] and 14[C] guanidinium ions as tracers. FlashPlate & Cytostar-Tplates in living cells avoids separation steps and are suitable for HTS.Scintillation plate technology has also advanced this method to HTSsuitability. Because of the functional aspects of the assay, theinformation content is reasonably good.

Yet another format measures the redistribution of membrane potentialusing the FLIPR system membrane potential kit (HTS) available fromMolecular Dynamics (a division of Amersham Biosciences, Piscataway,N.J.). This method is limited to slow membrane potential changes. Someproblems may result from the fluorescent background of compounds. Testcompounds may also directly influence the fluidity of the cell membraneand lead to an increase in intracellular dye concentrations. Still,because of the functional aspects of the assay, the information contentis reasonably good.

Sodium dyes can be used to measure the rate or amount of sodium ioninflux through a channel. This type of assay provides a very highinformation content regarding potential channel blockers. The assay isfunctional and would measure Na+ influx directly. CoroNa Red, SBFIand/or sodium green (Molecular Probes, Inc. Eugene Oreg.) can be used tomeasure Na influx; all are Na responsive dyes. They can be used incombination with the FLIPR instrument. The use of these dyes in a screenhas not been previously described in the literature. Calcium dyes mayalso have potential in this format.

In another assay, FRET based voltage sensors are used to measure theability of a test compound to directly block Na influx. Commerciallyavailable HTS systems include the VIPR™ II FRET system (LifeTechnologies, or Aurora Biosciences Corporation, San Diego, Calif., adivision of Vertex Pharmaceuticals, Inc.) which may be used inconjunction with FRET dyes, also available from Aurora Biosciences. Thisassay measures sub-second responses to voltage changes. There is norequirement for a modifier of channel function. The assay measuresdepolarization and hyperpolarizations, and provides ratiometric outputsfor quantification. A somewhat less expensive MTS version of this assayemploys the FLEXstation™ (Molecular Devices Corporation) in conjunctionwith FRET dyes from Aurora Biosciences. Other methods of testing thecompounds disclosed herein are also readily known and available to thoseskilled in the art.

Modulating agents so identified are then tested in a variety of in vivomodels so as to determine if they alleviate pain, especially chronicpain or other conditions such as cancer and pruritus (itch) with minimaladverse events. The assays described below in the Biological AssaysSection are useful in assessing the biological activity of the instantcompounds.

Typically, the efficacy of a compound of the invention is expressed byits IC50 value (“Inhibitory Concentration—50%”), which is the measure ofthe amount of compound required to achieve 50% inhibition of theactivity of the target sodium channel over a specific time period. Forexample, representative compounds of the present invention havedemonstrated IC50's ranging from less than 100 nanomolar to less than 10micromolar in the patch voltage clamp NaV1.7 electrophysiology assaydescribed herein.

In another aspect of the invention, the compounds of the invention canbe used in in vitro or in vivo studies as exemplary agents forcomparative purposes to find other compounds also useful in treatmentof, or protection from, the various diseases disclosed herein.

Another aspect of the invention relates to inhibiting NaV1.1, NaV1.2,NaV1.3, NaV1.4, NaV1.5, NaV1.6, NaV1.7, NaV1.8, or NaV1.9 activity,preferably NaV1.7 activity, in a biological sample or a mammal,preferably a human, which method comprises administering to the mammal,preferably a human, or contacting said biological sample with a compoundas described herein or a pharmaceutical composition comprising acompound as described herein. The term “biological sample”, as usedherein, includes, without limitation, cell cultures or extracts thereof;biopsied material obtained from a mammal or extracts thereof, and blood,saliva, urine, feces, semen, tears, or other body fluids or extractsthereof.

Inhibition of NaV1.1, NaV1.2, NaV1.3, NaV1.4, NaV1.5, NaV1.6, NaV1.7,NaV1.8, or NaV1.9 activity in a biological sample is useful for avariety of purposes that are known to one of skill in the art. Examplesof such purposes include, but are not limited to, the study of sodiumion channels in biological and pathological phenomena; and thecomparative evaluation of new sodium ion channel inhibitors.

The compounds of the invention (or stereoisomers, geometric isomers,tautomers, solvates, metabolites, isotopes, pharmaceutically acceptablesalts, or prodrugs thereof) and/or the pharmaceutical compositionsdescribed herein which comprise a pharmaceutically acceptable excipientand one or more compounds of the invention, can be used in thepreparation of a medicament for the treatment of sodium channel-mediateddisease or condition in a mammal.

Combination Therapy

The compounds of the invention may be usefully combined with one or moreother compounds of the invention or one or more other therapeutic agentor as any combination thereof, in the treatment of sodiumchannel-mediated diseases and conditions. For example, a compound of theinvention may be administered simultaneously, sequentially or separatelyin combination with other therapeutic agents, including, but not limitedto:

opiates analgesics, e.g., morphine, heroin, cocaine, oxymorphine,levorphanol, levallorphan, oxycodone, codeine, dihydrocodeine,propoxyphene, nalmefene, fentanyl, hydrocodone, hydromorphone,meripidine, methadone, nalorphine, naloxone, naltrexone, buprenorphine,butorphanol, nalbuphine and pentazocine;

non-opiate analgesics, e.g., acetomeniphen, salicylates (e.g., aspirin);

nonsteroidal antiinflammatory drugs (NSAIDs), e.g., ibuprofen, naproxen,fenoprofen, ketoprofen, celecoxib, diclofenac, diflusinal, etodolac,fenbufen, fenoprofen, flufenisal, flurbiprofen, ibuprofen, indomethacin,ketoprofen, ketorolac, meclofenamic acid, mefenamic acid, meloxicam,nabumetone, naproxen, nimesulide, nitroflurbiprofen, olsalazine,oxaprozin, phenylbutazone, piroxicam, sulfasalazine, sulindac, tolmetinand zomepirac;

anticonvulsants, e.g., carbamazepine, oxcarbazepine, lamotrigine,valproate, topiramate, gabapentin and pregabalin;

antidepressants such as tricyclic antidepressants, e.g., amitriptyline,clomipramine, despramine, imipramine and nortriptyline;

COX-2 selective inhibitors, e.g., celecoxib, rofecoxib, parecoxib,valdecoxib, deracoxib, etoricoxib, and lumiracoxib;

alpha-adrenergics, e.g., doxazosin, tamsulosin, clonidine, guanfacine,dexmetatomidine, modafinil, and 4-amino-6,7-dimethoxy-2-(5-methanesulfonamido-1,2,3,4-tetrahydroisoquinol-2-yl)-5-(2-pyridyl) quinazoline;

barbiturate sedatives, e.g., amobarbital, aprobarbital, butabarbital,butabital, mephobarbital, metharbital, methohexital, pentobarbital,phenobartital, secobarbital, talbutal, theamylal and thiopental;tachykinin (NK) antagonist, particularly an NK-3, NK-2 or NK-1antagonist, e.g., (aR,9R)-7-[3,5-bis(trifluoromethyl)benzyl)]-8,9,10,11-tetrahydro-9-methyl-5-(4-methylphenyl)-7H-[1,4]diazocino[2,1-g][1,7]-naphthyridine-6-13-dione(TAK-637),5-[[2R,3S)-2-[(1R)-1-[3,5-bis(trifluoromethylphenyl]ethoxy-3-(4-fluorophenyl)-4-morpholinyl]-methyl]-1,2-dihydro-3H-1,2,4-triazol-3-one(MK-869), aprepitant, lanepitant, dapitant or3-[[2-methoxy5-(trifluoromethoxy)phenyl]-methylamino]-2-phenylpiperidine(2S,3S);

coal-tar analgesics, in particular paracetamol;

serotonin reuptake inhibitors, e.g., paroxetine, sertraline,norfluoxetine (fluoxetine desmethyl metabolite), metabolitedemethylsertraline, ′3 fluvoxamine, paroxetine, citalopram, citaloprammetabolite desmethylcitalopram, escitalopram, d,l-fenfluramine,femoxetine, ifoxetine, cyanodothiepin, litoxetine, dapoxetine,nefazodone, cericlamine, trazodone and fluoxetine;

noradrenaline (norepinephrine) reuptake inhibitors, e.g., maprotiline,lofepramine, mirtazepine, oxaprotiline, fezolamine, tomoxetine,mianserin, buproprion, buproprion metabolite hydroxybuproprion,nomifensine and viloxazine (Vivalan®)), especially a selectivenoradrenaline reuptake inhibitor such as reboxetine, in particular(S,S)-reboxetine, and venlafaxine duloxetine neurolepticssedative/anxiolytics;

dual serotonin-noradrenaline reuptake inhibitors, such as venlafaxine,venlafaxine metabolite O-desmethylvenlafaxine, clomipramine,clomipramine metabolite desmethylclomipramine, duloxetine, milnacipranand imipramine;

acetylcholinesterase inhibitors such as donepezil;

5-HT3 antagonists such as ondansetron;

metabotropic glutamate receptor (mGluR) antagonists;

local anaesthetic such as mexiletine and lidocaine;

corticosteroid such as dexamethasone;

antiarrhythimics, e.g., mexiletine and phenytoin;

muscarinic antagonists, e.g., tolterodine, propiverine, tropsium tchloride, darifenacin, solifenacin, temiverine and ipratropium;

cannabinoids;

vanilloid receptor agonists (e.g., resinferatoxin) or antagonists (e.g.,capsazepine);

sedatives, e.g., glutethimide, meprobamate, methaqualone, anddichloralphenazone;

anxiolytics such as benzodiazepines,

antidepressants such as mirtazapine,

topical agents (e.g., lidocaine, capsacin and resiniferotoxin);

muscle relaxants such as benzodiazepines, baclofen, carisoprodol,chlorzoxazone, cyclobenzaprine, methocarbamol and orphrenadine;

anti-histamines or H1 antagonists;

NMDA receptor antagonists;

5-HT receptor agonists/antagonists;

PDEV inhibitors;

Tramadol®;

cholinergic (nicotinc) analgesics;

alpha-2-delta ligands;

prostaglandin E2 subtype antagonists;

leukotriene B4 antagonists;

5-lipoxygenase inhibitors; and

5-HT3 antagonists.

Sodium channel-mediated diseases and conditions that may be treatedand/or prevented using such combinations include but not limited to,pain, central and peripherally mediated, acute, chronic, neuropathic aswell as other diseases with associated pain and other central nervousdisorders such as epilepsy, anxiety, depression and bipolar disease; orcardiovascular disorders such as arrhythmias, atrial fibrillation andventricular fibrillation; neuromuscular disorders such as restless legsyndrome and muscle paralysis or tetanus; neuroprotection againststroke, neural trauma and multiple sclerosis; and channelopathies suchas erythromyalgia and familial rectal pain syndrome.

As used herein “combination” refers to any mixture or permutation of oneor more compounds of the invention and one or more other compounds ofthe invention or one or more additional therapeutic agent. Unless thecontext makes clear otherwise, “combination” may include simultaneous orsequentially delivery of a compound of the invention with one or moretherapeutic agents. Unless the context makes clear otherwise,“combination” may include dosage forms of a compound of the inventionwith another therapeutic agent. Unless the context makes clearotherwise, “combination” may include routes of administration of acompound of the invention with another therapeutic agent. Unless thecontext makes clear otherwise, “combination” may include formulations ofa compound of the invention with another therapeutic agent. Dosageforms, routes of administration and pharmaceutical compositions include,but are not limited to, those described herein.

The invention will be more fully understood by reference to thefollowing examples. They should not, however, be construed as limitingthe scope of the invention.

EXAMPLES

These examples serve to provide guidance to a skilled artisan to prepareand use the compounds, compositions and methods of the invention. Whilea particular embodiment of the present invention are described, theskilled artisan will appreciate that various changes and modificationscan be made without departing from the spirit and scope of theinventions.

The chemical reactions in the examples described can be readily adaptedto prepare a number of other compounds of the invention, and alternativemethods for preparing the compounds of this invention are deemed to bewithin the scope of this invention. For example, the synthesis ofnon-exemplified compounds according to the invention can be successfullyperformed by modifications apparent to those skilled in the art, forexample, by appropriately protecting interfering groups by utilizingother suitable reagents known in the art other than those described,and/or by making routine modifications of reaction conditions.

In the examples below, unless otherwise indicated all temperatures areset forth in degrees Celsius. Commercially available reagents werepurchased from suppliers such as Aldrich Chemical Company, Lancaster,TCI or Maybridge and were used without further purification unlessotherwise indicated. The reactions set forth below were done under apositive pressure of nitrogen or argon or with a drying tube (unlessotherwise stated) in anhydrous solvents, and the reaction flasks weretypically fitted with rubber septa for the introduction of substratesand reagents via syringe. Glassware was oven dried and/or heat dried. ¹HNMR spectra were obtained in deuterated CDCl₃, d₆-DMSO, CH₃OD ord₆-acetone solvent solutions (reported in ppm) using or trimethylsilane(TMS) or residual non-deuterated solvent peaks as the referencestandard. When peak multiplicities are reported, the followingabbreviates are used: s (singlet), d (doublet), t (triplet), q(quartet), m (multiplet), br (broad), dd (doublet of doublets), dt(doublet of triplets). Coupling constants, when given, are reported inHz (Hertz).

All abbreviations used to describe reagents, reaction conditions orequipment are intended to be consistent with the definitions set forthin the “List of standard abbreviates and acronyms”. The chemical namesof discrete compounds of the invention were obtained using the structurenaming feature of ChemDraw naming program.

Abbreviations

MeCN Acetonitrile

EtOAc Ethyl acetate

DCE Dichloroethane

DCM Dichloromethane

DIPEA Diisopropylethylamine

DEA Diethylamine

DMAP 4-dimethylaminopyridine

DMF N,N-Dimethylformamide

DMSO Dimethyl sulfoxide

FA Formic acid

IPA Isopropyl alcohol

TFA Trifluoroacetic acid

EDCI 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride

HCl Hydrochloric acid

HPLC High Pressure Liquid Chromatography

LCMS Liquid Chromatography Mass Spectrometry

MeOH Methanol

NMP N-methyl-2-pyrrolidone

RPHPLC Reverse phase high pressure liquid chromatography

RT Retention time

SFC Supercritical Fluid Chromatography

THF Tetrahydrofuran

TEA Triethylamine

Example 1 & Example 2

(4aR,8aR)-1-(2,4′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2,4′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Step 1:

Rac-trans-tert-butyl4-((4-bromo-5-fluoro-2-methoxyphenyl)amino)-3-hydroxypiperidine-1-carboxylate

To a solution of 4-bromo-5-fluoro-2-methoxyaniline (1.33 g, 6.02 mmol),tert-butyl 7-oxa-3-azabicyclo[4.1.0]heptane-3-carboxylate (1.0 g, 5.02mmol) in 1,2-dichloroethane (30 mL) was added Sc(OTf)₃ (371 mg, 0.75mmol). The mixture was stirred at room temperature for 16 h. The mixturewas concentrated in vacuo and the crude residue was purified by silicagel chromatography (solvent gradient: 0-25% EtOAc in petroleum ether) togive the title compound (1.1 g, 52%) as yellow oil. ¹H NMR (400 MHz,CDCl₃) δ 6.82 (d, J=6.4 Hz, 1H), 6.54 (d, J=10.4 Hz, 1H), 4.28-4.24 (m,1H), 4.06-4.00 (m, 1H), 3.82 (s, 3H), 3.54-3.46 (m, 1H), 3.24-3.19 (m,1H), 2.75-2.88 (m, 2H), 2.09-2.05 (m, 1H), 1.45 (s, 9H), 1.38-1.31 (m,1H).

Rac-trans-tert-butyl1-(4-bromo-5-fluoro-2-methoxyphenyl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-carboxylate

To a solution of rac-trans-tert-butyl4-((4-bromo-5-fluoro-2-methoxyphenyl)amino)-3-hydroxypiperidine-1-carboxylate(1.1 g, 2.62 mmol) in THF (12 mL) was added NaH (260 mg, 6.56 mmol) at0° C. under a nitrogen atmosphere. After stirring at 0° C. for 0.5 h,ethyl 2-bromoacetate (0.46 mL, 4.2 mmol) was added dropwise. The mixturewas warmed to room temperature and stirred for an additional 16 h. Thereaction was quenched with sat. aq. NH₄Cl. The mixture was concentratedin vacuo and the residue was purified by silica gel chromatography(solvent gradient: 0-20% EtOAc in petroleum ether) to give the titlecompound (1.0 g, 83%) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ7.14-7.11 (m, 1H), 6.97-6.92 (m, 1H), 4.52-4.39 (m, 3H), 4.20-4.10 (m,1H), 3.84-3.80 (m, 3H), 3.78-3.71 (m, 1H), 3.58-3.51 (m, 1H), 2.73-2.55(m, 2H), 1.46 (s, 9H), 1.40-1.28 (m, 2H) Step 3:

Rac-trans-1-(4-bromo-5-fluoro-2-methoxyphenyl)hexahydro-1H-pyrido[3,4-b][1,4]oxazin-2(3H)-one

To a stirred solution of rac-(trans)-tert-butyl1-(4-bromo-5-fluoro-2-methoxyphenyl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-carboxylate(260 mg, 0.57 mmol) in EtOAc (5 mL) was added 4 M HCl (0.71 mL, 2.83mmol) in EtOAc. The mixture was stirred at room temperature for 2 h. Thereaction was quenched with sat. aq. NaHCO₃ (10 mL) and extracted withEtOAc (10 mL×2). The combined organic layers were dried over anhydrousNa₂SO₄, filtered and concentrated in vacuo to give the title compound(210 mg, crude) as yellow oil that required no further purification. ¹HNMR (400 MHz, DMSO-d₆) δ 7.49-7.40 (m, 1H), 7.27-7.24 (m, 1H), 4.32-4.21(m, 2H), 3.85-3.73 (m, 3H), 3.57-3.46 (m, 2H), 3.11-3.01 (m, 1H),2.85-2.72 (m, 1H), 2.42-2.22 (m, 2H), 1.33-1.15 (m, 2H).

Step 4:

Rac-trans-1-(3′-bromo-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

To a solution of imidazole (1.99 g, 29.23 mmol) and 3-aminoisoxazole(787 mg, 9.35 mmol) in DCM (4 mL) was added SO₂Cl₂ (0.77 mL, 9.35 mmol)at −78° C. Then the mixture was warmed to room temperature and stirredfor 0.5 h. A solution ofrac-trans-1-(4-bromo-5-fluoro-2-methoxyphenyl)hexahydro-1H-pyrido[3,4-b][1,4]oxazin-2(3H)-one(210 mg, 0.58 mmol) in DCM (4 mL) was added dropwise. The reactionmixture was heated to 40° C. for 1 h. The reaction mixture was quenchedwith water (20 mL) and extracted with DCM (20 mL). The organic layer waswashed with brine (5 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated in vacuo. The crude residue was purified by silica gelchromatography (solvent gradient: 0-5% methanol in DCM) to give thetitle compound (260 mg, 88%) as yellow solid. ¹H NMR (400 MHz, CDCl₃) δ8.23 (s, 1H), 8.08-8.06 (m, 1H), 7.14-7.10 (m, 1H), 6.95-6.90 (m, 1H),6.42-6.38 (m, 1H), 4.49-4.33 (m, 2H), 3.84-3.79 (m, 3H), 3.76-3.59 (m,4H), 2.82-2.67 (m, 2H), 1.54-1.35 (m, 2H).

Rac-trans-1-(2,4′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

To a solution ofrac-trans-1-(3′-bromo-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(300 mg, 0.42 mmol),bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(II)(59 mg, 0.08 mmol), Cs₂CO₃ (258 mg, 0.79 mmol) and CuCl (123 mg, 1.25mmol) in DMF (5 mL) was added(4-fluoro-3-(trifluoromethyl)phenyl)boronic acid (259 mg, 1.25 mmol).The reaction mixture was heated to 50° C. for 2 h under a nitrogenatmosphere. After cooling to room temperature, the reaction mixture wasquenched with water (50 mL), extracted with EtOAc (50 mL). The organiclayer was washed with brine (20 mL), dried over anhydrous Na₂SO₄,filtered and concentrated in vacuo. The residue was purified by reversephase chromatography (acetonitrile 65-95/0.225% formic acid in water) togive the title compound (90 mg, 37%) as yellow oil. LCMS (ESI) m/z:589.0 [M+H]⁺.

Step 6:

(4aR,8aR)-1-(2,4′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2,4′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Rac-trans-1-(2,4′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamidewas separated by using chiral SFC (Phenomenex C2 (250 mm*30 mm, 10 um),Supercritical CO₂/EtOH+0.1% NH₄OH=60/40; 50 mL/min) to give(4aR,8aR)-1-(2,4′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(11 mg, first peak) as white powder and(4aS,8aS)-1-(2,4′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(11 mg, second peak) as a white solid. Absolute configuration wasarbitrarily assigned to each enantiomer. Example 1: LCMS (ESI) m/z:589.1 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.73-8.66 (m, 1H), 8.04-7.94(m, 2H), 7.71-7.63 (m, 1H), 7.42-7.19 (m, 2H), 6.37 (s, 1H), 4.41-4.27(m, 2H), 3.88-3.78 (m, 4H), 3.76-3.53 (m, 3H), 2.83-2.69 (m, 2H),1.42-1.35 (m, 1H), 1.41-1.23 (m, 2H). Example 2: LCMS (ESI) m/z: 589.1[M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.74-8.65 (m, 1H), 8.03-7.92 (m,2H), 7.71-7.63 (m, 1H), 7.43-7.18 (m, 2H), 6.37 (s, 1H), 4.41-4.27 (m,2H), 3.89-3.78 (m, 4H), 3.76-3.53 (m, 3H), 2.83-2.68 (m, 2H), 1.42-1.19(m, 2H)

Example 3 & Example 4

(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-bi][1,4]oxazine-6(7H)-sulfonamide

Step 1:

Rac-trans-tert-butyl1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-carboxylate

To a solution of 3-(trifluoromethyl)phenylboronic acid (11.83 g, 62.27mmol), rac-trans-tert-butyl1-(4-bromo-5-fluoro-2-methoxyphenyl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-carboxylate(26.0 g, 56.61 mmol) and K₂CO₃ (23.47 g, 169.82 mmol) in water (50 mL)and 1,4-dioxane (260 mL) was added tetrakis(triphenylphosphine)palladium(6.54 g, 5.66 mmol). The reaction mixture was heated to 100° C. for 16 hunder a nitrogen atmosphere. After cooling to room temperature, thereaction mixture was diluted with water (50 mL) and extracted with EtOAc(150 mL×3). The combined organic layers were washed with water (160 mL),brine (140 mL), dried over anhydrous Na₂SO₄, filtered and concentratedin vacuo. The crude residue was purified by silica gel chromatography(solvent gradient: 0-35% ethyl acetate in petroleum ether) to give thetitle compound (29 g, 98%) as yellow oil. ¹H NMR (400 MHz, CDCl₃) δ 7.77(s, 1H), 7.75-7.64 (m, 2H), 7.62-7.56 (m, 1H), 7.05-6.94 (m, 2H),4.59-4.30 (m, 3H), 4.26-4.00 (m, 1H), 3.92-3.85 (m, 3H), 3.84-3.78 (m,1H), 3.67-3.52 (m, 1H), 2.85-2.54 (m, 2H), 1.68-1.57 (m, 1H), 1.48 (s,9H), 1.40-1.30 (m, 1H).

Step 2:

Rac-trans-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazin-2(3H)-one

To a solution of Rac-trans-tert-butyl1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-carboxylate(29.0 g, 55.29 mmol) in EtOAc (260 mL) was added 4M HCl (276 mL, 1105.8mmol) in EtOAc. The reaction mixture was stirred at room temperature for2 h. The reaction mixture was concentrated in vacuo to give the titlecompound (23 g, 98%) as a yellow solid that required no furtherpurification. ¹H NMR (400 MHz, CDCl₃) δ 8.25-8.20 (m, 1H), 7.78-7.75 (m,1H), 7.70-7.62 (m, 2H), 7.60-7.50 (m, 1H), 6.99-6.94 (m, 2H), 4.55-4.41(m, 2H), 3.91-3.85 (m, 3H), 3.83-3.54 (m, 2H), 3.39-3.31 (m, 1H),3.09-2.98 (m, 1H), 2.71-2.51 (m, 2H), 1.62-1.24 (m, 2H).

Step 3:

Rac-trans-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

To a solution of imidazole (18.45 g, 270.98 mmol) and 3-aminoisoxazole(7.29 g, 86.71 mmol) in DCM (500 mL) was added SO₂Cl₂ (11.70 g, 86.71mmol) at −78° C. The reaction mixture was stirred at room temperaturefor 30 min. Thenrac-trans-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazin-2(3H)-one(23.0 g, 54.2 mmol) in DCM (500 mL) was added dropwise. The reactionmixture was heated to 40° C. for 1 h. The reaction mixture was dilutedwith water (50 mL) and extracted with EtOAc (200 mL). The organic layerwas washed with water (50 mL), sat. aq. citric acid (50 mL), brine (40mL), dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo.The residue was purified by silica gel chromatography (solvent gradient:0-25% EtOAc (10% MeOH) in petroleum ether) to give the title compound(22 g, crude). The crude product (28 g) were recrystallized frompropan-2-ol (90 mL) to give the title compound (18.7 g, 60%) as yellowsolid. LCMS (ESI) m/z: 571.1 [M+H]⁺.

Step 4:

(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Rac-trans-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(18.7 g) was separated by using chiral SFC (Chiralpak AD (250 mm*50 mm,10 um)), Supercritical CO₂/EtOH+0.1% NH₄OH=60/40; 200 mL/min) to give(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(5.92 g, 31.7%, second peak) as a white solid and(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(7.93 g, 42%, first peak) as a white solid. Absolute configuration wasarbitrarily assigned to each enantiomer. Example 3: LCMS (ESI) m/z:571.2 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 11.20 (s, 1H), 8.81-8.76 (m,1H), 7.95-7.90 (m, 2H), 7.84-7.72 (m, 2H), 7.42-7.19 (m, 2H), 6.41-6.38(m, 1H), 4.41-4.27 (m, 2H), 3.90-3.79 (m, 4H), 3.73-3.56 (m, 3H),2.88-2.65 (m, 2H), 1.53-1.18 (m, 2H). Example 4: LCMS (ESI) m/z: 571.2[M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 11.20 (s, 1H), 8.81-8.76 (m, 1H),7.95-7.90 (m, 2H), 7.84-7.72 (m, 2H), 7.42-7.19 (m, 2H), 6.41-6.38 (m,1H), 4.41-4.27 (m, 2H), 3.90-3.79 (m, 4H), 3.73-3.56 (m, 3H), 2.88-2.65(m, 2H), 1.53-1.18 (m, 2H).

Example 5 & Example 6

(4aR,8aR)-1-(2,3′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2,3′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Step 1:

Rac-trans-1-(2,3′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Following the procedure described in Example 1, step 5 and makingnon-critical variations as required to replace(4-fluoro-3-(trifluoromethyl)phenyl)boronic acid with(3-fluoro-5-(trifluoromethyl)phenyl)boronic acid,Rac-trans-1-(2,3′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(150 mg, 43% yield) was obtained as yellow oil.

Step 2:

(4aR,8aR)-1-(2,3′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2,3′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Rac-trans-(4aR,8aR)-1-(2,3′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(150 mg) was separated by using chiral SFC (chiralpak IC (250 mm*30 mm,5 um), Supercritical CO₂/EtOH+0.1% NH₄OH=55/45; 60 mL/min) to give(4aR,8aR)-1-(2,3′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(15 mg, first peak) as a white solid and(4aS,8aS)-1-(2,3′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(16 mg, second peak) as a white solid. Absolute configuration wasarbitrarily assigned to each enantiomer. Example 5: LCMS (ESI) m/z:589.1 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 11.18 (s, 1H), 8.78-8.73 (m,1H), 7.86 (d, J=8.8 Hz, 1H), 7.83-7.75 (m, 2H), 7.45-7.20 (m, 2H),6.41-6.37 (m, 1H), 4.45-4.26 (m, 2H), 3.90-3.72 (m, 5H), 3.71-3.58 (m,2H), 2.87-2.69 (m, 2H), 1.53-1.20 (m, 2H). Example 6: LCMS (ESI) m/z:589.1 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 11.18 (s, 1H), 8.80-8.71 (m,1H), 7.87 (d, J⁼8.8 Hz, 1H), 7.83-7.76 (m, 2H), 7.45-7.21 (m, 2H),6.41-6.37 (m, 1H), 4.42-4.28 (m, 2H), 3.91-3.71 (m, 5H), 3.70-3.56 (m,2H), 2.86-2.69 (m, 2H), 1.51-1.20 (m, 2H).

Example 7 & Example 8

(4aR,8aR)-1-(2,2′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2,2′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Step 1:

Rac-trans-1-(2,2′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Following the procedure described in Example 1, step 5 and makingnon-critical variations as required to replace(4-fluoro-3-(trifluoromethyl)phenyl)boronic acid with(2-fluoro-5-(trifluoromethyl)phenyl)boronic acid,Rac-trans-1-(2,2′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(120 mg, 29%) as yellow oil.

(4aR,8aR)-1-(2,2′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2,2′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Rac-trans-1-(2,2′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(120 mg) was separated by using chiral SFC (chiralpak IC (250 mm*30 mm,5 um), Supercritical CO₂/EtOH+0.1% NH₄OH=60/40; 50 mL/min) to give(4aR,8aR)-1-(2,2′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(32 mg, first peak) as a white solid and(4aS,8aS)-1-(2,2′-difluoro-5-methoxy-5′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(30 mg, second peak) as a white solid. Absolute configuration wasarbitrarily assigned to each enantiomer. Example 7: LCMS (ESI) m/z:589.1 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 11.15 (s, 1H), 8.70-8.61 (m,1H), 7.96-7.88 (m, 2H), 7.62-7.57 (m, 1H), 7.39-7.18 (m, 2H), 6.33 (s,1H), 4.37-4.30 (m, 2H), 3.82-3.74 (m, 4H), 3.70-3.48 (m, 3H), 2.79-2.60(m, 2H), 1.47-1.32 (m, 2H). Example 8: LCMS (ESI) m/z: 589.1 [M+H]⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 11.15 (s, 1H), 8.76-8.69 (m, 1H), 7.96-7.88 (m,2H), 7.62-7.57 (m, 1H), 7.39-7.18 (m, 2H), 6.37-6.34 (m, 1H), 4.38-4.28(m, 2H), 3.82-3.74 (m, 4H), 3.72-3.50 (m, 3H), 2.82-2.69 (m, 2H),1.49-1.34 (m, 2H).

Example 9 & Example 10

(4aR,8aR)-1-(2,3′-difluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2,3′-difluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Step 1:

Rac-trans-1-(2,3′-difluoro-5-methoxy-[1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Following the procedure described in Example 1, step 5 and makingnon-critical variations as required to replace(4-fluoro-3-(trifluoromethyl)phenyl)boronic acid with(3-fluorophenyl)boronic acid,Rac-trans-1-(2,3′-difluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(60 mg, 58%) was obtained as yellow oil. LCMS (ESI) m/z: 521.1 [M+H]⁺.

(4aR,8aR)-1-(2,3′-difluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2,3′-difluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Rac-trans-(4aR,8aR)-1-(2,3′-difluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(150 mg) was separated by using chiral SFC (Phenomenex C2 (250 mm*30 mm,5 um), Supercritical CO₂/EtOH+0.1% NH₄OH=50/50; 80 mL/min) to give(4aR,8aR)-1-(2,3′-difluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(28 mg, first peak) as a white solid and(4aS,8aS)-1-(2,3′-difluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(29 mg, second peak) as a white solid. Absolute configuration wasarbitrarily assigned to each enantiomer. Example 9: LCMS (ESI) m/z:521.0 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 11.18 (s, 1H), 8.81-8.74 (m,1H), 7.59-7.43 (m, 3H), 7.39-7.15 (m, 3H), 6.41-6.36 (m, 1H), 4.42-4.27(m, 2H), 3.89-3.78 (m, 4H), 3.75-3.56 (m, 3H), 2.88-2.68 (m, 2H),1.54-1.17 (m, 2H). Example 10: LCMS (ESI) m/z: 521.0 [M+H]⁺. ¹H NMR (400MHz, DMSO-d₆) δ 11.19 (s, 1H), 8.81-8.73 (m, 1H), 7.59-7.42 (m, 3H),7.39-7.15 (m, 3H), 6.41-6.36 (m, 1H), 4.42-4.26 (m, 2H), 3.90-3.78 (m,4H), 3.77-3.56 (m, 3H), 2.88-2.70 (m, 2H), 1.53-1.18 (m, 2H).

Example 11 & Example 12

(4aR,8aR)-1-(2,2′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2,2′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Step 1:

Rac-trans-(4aR,8aR)-1-(2,2′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Following the procedure described in Example 1, step 5 and makingnon-critical variations as required to replace(4-fluoro-3-(trifluoromethyl)phenyl)boronic acid with(2-fluoro-3-(trifluoromethyl)phenyl)boronic acid,Rac-trans-1-(2,2′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(40 mg, 9%) was obtained as yellow oil. LCMS (ESI) m/z: 589.1 [M+H]⁺.

Step 2:

(4aR,8aR)-1-(2,2′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2,2′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Rac-trans-1-(2,2′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(40 mg) was separated by using chiral SFC (Chiralpak C2 (250 mm*30 mm, 5um), Supercritical CO₂/EtOH+0.1% NH₄OH=40/60; 55 mL/min) to give(4aR,8aR)-1-(2,2′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(12 mg, first peak) as white powder and(4aS,8aS)-1-(2,2′-difluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(12 mg, second peak) as white powder. Absolute configuration wasarbitrarily assigned to each enantiomer. Example 11: LCMS (ESI) m/z:589.2 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.36 (s, 1H), 7.90-7.85 (m,2H), 7.56-7.51 (m, 1H), 7.23-7.17 (m, 2H), 6.24 (s, 1H), 4.36-4.23 (m,2H), 3.83-3.74 (m, 3H), 3.73-3.51 (m, 2H), 3.48-3.36 (m, 2H), 2.60-2.50(m, 2H), 1.45-1.20 (m, 2H). Example 12: LCMS (ESI) m/z: 589.2 [M+H]⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 8.39 (s, 1H), 7.92-7.83 (m, 2H), 7.55-7.52 (m,1H), 7.25-7.15 (m, 2H), 6.25 (s, 1H), 4.37-4.22 (m, 2H), 3.80-3.74 (m,3H), 3.70-3.52 (m, 2H), 3.50-3.40 (m, 2H), 2.60-2.50 (m, 2H), 1.40-1.20(m, 2H).

Example 13 & Example 14

(4aR,8aR)-1-(3′-chloro-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(3′-chloro-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Step 1:

Rac-trans-1-(3′-chloro-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Following the procedure described in Example 1, step 5 and makingnon-critical variations as required to replace(4-fluoro-3-(trifluoromethyl)phenyl)boronic acid with(3-chlorophenyl)boronic acid,Rac-trans-1-(3′-chloro-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(50 mg, 23%) was obtained as a white solid. LCMS (ESI) m/z: 537.1[M+H]⁺.

Step 2:

(4aR,8aR)-1-(3′-chloro-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(3′-chloro-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Rac-trans-1-(3′-chloro-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(50 mg) was separated by using chiral SFC (Chiralpak C2 IC (250 mm*30mm, 5 um), Supercritical CO₂/EtOH+0.1% NH₄OH=40/60; 55 mL/min) to give(4aR,8aR)-1-(3′-chloro-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(4 mg, first peak) as a white solid and(4aS,8aS)-1-(3′-chloro-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(7 mg, second peak) as a white solid. Absolute configuration wasarbitrarily assigned to each enantiomer. Example 13: LCMS (ESI) m/z:537.0 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.32 (s, 1H), 7.70-7.67 (m,1H), 7.57-7.51 (m, 2H), 7.24-7.19 (m, 2H), 6.25 (s, 1H), 4.44-4.34 (m,2H), 3.87-3.80 (m, 3H), 3.77-3.67 (m, 1H), 3.67-3.40 (m, 3H), 2.54-2.50(m, 2H), 1.45-1.20 (m, 2H). Example 14: LCMS (ESI) m/z: 537.2 [M+H]⁺. HNMR (400 MHz, DMSO-d₆) δ 8.18 (s, 1H), 7.68-7.63 (m, 1H), 7.57-7.52 (m,2H), 7.48-7.46 (m, 1H), 7.20-7.15 (m, 2H), 6.19-6.17 (m, 1H), 4.36-4.21(m, 2H), 3.83-3.76 (m, 3H), 3.67-3.56 (m, 2H), 3.53-3.39 (m, 2H),2.35-2.31 (m, 2H), 1.42-1.20 (m, 2H).

Example 15 & Example 16

(4aR,8aR)-1-(3′-(difluoromethyl)-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(3′-(difluoromethyl)-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Step 1:

Following the procedure described in Example 1, step 5 and makingnon-critical variations as required to replace(4-fluoro-3-(trifluoromethyl)phenyl)boronic acid with(3-(difluoromethyl)-phenyl)boronic acid,rac-trans-1-(3′-(difluoromethyl)-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(30 mg, 14%) was obtained as a white solid.

Step 2:

(4aR,8aR)-1-(3′-(difluoromethyl)-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(3′-(difluoromethyl)-2-fluoro-5-methoxy-[1,1′-1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Rac-trans-1-(3′-(difluoromethyl)-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(30 mg) was separated by using chiral SFC (Chiralpak C2 IC (250 mm*30mm, 5 um), Supercritical CO₂/EtOH+0.1% NH₄OH=45/55; 50 mL/min) to give(4aR,8aR)-1-(3′-(difluoromethyl)-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(6 mg, first peak) as a white solid and(4aS,8aS)-1-(3′-(difluoromethyl)-2-fluoro-5-methoxy-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(8.5 mg, second peak) as a white solid. Absolute configuration wasarbitrarily assigned to each enantiomer. Example 15: LCMS (ESI) m/z:553.2 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.28 (s, 1H), 7.82-7.76 (m,2H), 7.69-7.65 (m, 2H), 7.45-7.22 (m, 2H), 7.20-7.00 (m, 1H), 6.24 (s,1H), 4.39-4.30 (m, 2H), 3.87-3.80 (m, 3H), 3.77-3.67 (m, 1H), 3.63-3.56(m, 1H), 3.47-3.40 (m, 3H), 1.46-1.33 (m, 2H), 1.32-1.23 (m, 1H).Example 16: LCMS (ESI) m/z: 553.2 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ8.30 (s, 1H), 7.82-7.75 (m, 2H), 7.69-7.61 (m, 2H), 7.45-7.22 (m, 2H),7.20-7.00 (m, 1H), 6.24 (s, 1H), 4.38-4.24 (m, 2H), 3.87-3.80 (s, 3H),3.72-3.70 (m, 1H), 3.65-3.56 (m, 1H), 3.45-3.42 (m, 1H), 3.29-3.26 (m,2H), 1.46-1.34 (m, 2H), 1.30-1.23 (s, 1H).

Example 17 & Example 18

(4aR,8aR)-N-(isoxazol-3-yl)-1-(3-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-N-(isoxazol-3-yl)-1-(3-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Step 1:

Rac-trans-1-(4-bromo-2-methoxyphenyl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Following the procedure described in Example 1, step 1-4 and makingnon-critical variations as required to replace4-bromo-5-fluoro-2-methoxyaniline with 4-bromo-2-methoxyaniline,Rac-trans-1-(4-bromo-2-methoxyphenyl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(110 mg, 25%) was obtained as a yellow solid. LCMS (ESI) m/z: 487.0[M+H]⁺.

Step 2:

Rac-trans-N-(isoxazol-3-yl)-1-(3-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Following the procedure described in Example 1, step 5 and makingnon-critical variations as required to replacerac-trans-1-(4-bromo-5-fluoro-2-methoxyphenyl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamidewithrac-trans-1-(4-bromo-2-methoxyphenyl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide,(4-fluoro-3-(trifluoromethyl)phenyl)boronic acid with(3-(trifluoromethyl)phenyl)boronic acid,rac-trans-N-(isoxazol-3-yl)-1-(3-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(95 mg, 76%) was obtained as a yellow solid. LCMS (ESI) m/z: 553.2[M+H].

Step 3:

(4aR,8aR)-N-(isoxazol-3-yl)-1-(3-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-N-(isoxazol-3-yl)-1-(3-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Rac-trans-(4aR,8aR)-N-(isoxazol-3-yl)-1-(3-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(90 mg) was separated by using chiral SFC (Chiralpak IC (250 mm*30 mm, 5um), Supercritical CO₂/EtOH+0.1% NH₄OH=50/50; 80 mL/min) to give(4aR,8aR)-N-(isoxazol-3-yl)-1-(3-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(20 mg, first peak) as a white solid and(4aS,8aS)-N-(isoxazol-3-yl)-1-(3-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(19 mg, second peak) as a white solid. Absolute configuration wasarbitrarily assigned to each enantiomer. Example 17: LCMS (ESI) m/z:553.1 [M+H]⁺. ¹H NMR (400 MHz, CD₃OD) δ 8.52 (s, 1H), 7.93-7.91 (m, 2H),7.69-7.63 (m, 2H), 7.33-7.20 (m, 3H), 6.44 (s, 1H), 4.48-4.38 (m, 2H),4.03-4.01 (m, 1H), 3.94-3.90 (m, 3H), 3.76-3.72 (m, 3H), 2.89-2.77 (m,2H), 1.44-1.48 (m, 1H), 1.38-1.24 (m, 1H). Example 18: LCMS (ESI) m/z:553.1 [M+H]⁺. ¹H NMR (400 MHz, CD₃OD) δ 8.65 (s, 1H), 7.94-7.86 (m, 2H),7.72-7.61 (m, 2H), 7.39-7.20 (m, 3H), 6.56 (s, 1H), 4.51-4.32 (m, 2H),4.05-4.00 (m, 1H), 3.97-3.88 (m, 3H), 3.80-3.65 (m, 3H), 2.95-2.70 (m,2H), 1.50-1.27 (m, 2H).

Example 19 & Example 20

(4aR,8aR)-1-(3-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(3-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamidede

Step 1:

Rac-trans-(4aR,8aR)-1-(4-bromo-2-fluoro-6-methoxyphenyl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Following the procedure described in Example 1, step 1-4 and makingnon-critical variations as required to replace4-bromo-5-fluoro-2-methoxyaniline with4-bromo-2-fluoro-6-methoxyaniline, Rac-trans-(4aR,8aR)-1-(4-bromo-2-fluoro-6-methoxyphenyl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(40 mg, 51%) was obtained as green oil. ¹H NMR (400 MHz, CDCl₃) δ 8.28(s, 1H), 7.07-6.88 (m, 2H), 6.49-6.44 (m, 1H), 4.51-4.39 (m, 2H),4.05-4.00 (m, 1H), 3.87-3.83 (m, 3H), 3.82-3.73 (m, 2H), 3.73-3.55 (m,2H), 3.50-3.45 (m, 1H), 2.85-2.72 (m, 2H), 1.59-1.43 (m, 2H).

Step 2:

Rac-trans-1-(3-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Following the procedure described in Example 1, step 5 and makingnon-critical variations as required to replaceRac-trans-1-(4-bromo-5-fluoro-2-methoxyphenyl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamidewithrac-trans-1-(4-bromo-2-fluoro-6-methoxyphenyl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide,(4-fluoro-3-(trifluoromethyl)phenyl)boronic acid with(3-(trifluoromethyl)phenyl)boronic acidRac-trans-1-(3-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(110 mg, 29% yield) was obtained as a white solid. LCMS (ESI) m/z: 571.1[M+H]⁺. H NMR (400 MHz, CDCl₃) δ 8.31 (s, 1H), 7.77 (s, 1H), 7.74-7.65(m, 2H), 7.62-7.56 (m, 1H), 7.02-6.98 (m, 1H), 6.93-6.90 (m, 1H), 6.48(s, 1H), 4.57-4.43 (m, 2H), 4.17-4.07 (m, 1H), 3.95-3.90 (m, 3H),3.85-3.65 (m, 3H), 2.88-2.77 (m, 2H), 2.06-2.04 (m, 1H), 1.29-1.24 (m,1H).

Step 3:

(4aR,8aR)-1-(3-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(3-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamidede

Rac-trans-1-(3-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(110 mg) was separated by using chiral SFC (Chiralpak AD (250 mm*30 mm,5 um), Supercritical CO₂/EtOH+0.1% NH₄OH=75/25; 60 mL/min) to give(4aR,8aR)-1-(3-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(30 mg, first peak) as a white solid and(4aS,8aS)-1-(3-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamidede(25 mg, second peak) as a white solid. Absolute configuration wasarbitrarily assigned to each enantiomer. Example 19: LCMS (ESI) m/z:571.1 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 8.31 (d, J=1.6 Hz, 1H), 7.78 (s,1H), 7.74-7.65 (m, 2H), 7.62-7.57 (m, 1H), 7.03-6.99 (m, 1H), 6.92 (s,1H), 6.48 (d, J=1.6 Hz, 1H), 4.56-4.45 (m, 2H), 4.10-4.05 (m, 1H), 3.94(s, 3H), 3.84-3.74 (m, 3H), 2.89-2.78 (m, 2H), 1.64-1.60 (m, 2H).Example 20: LCMS (ESI) m/z: 571.1 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ8.26-8.20 (m, 1H), 8.12-8.05 (m, 2H), 7.81-7.70 (m, 2H), 7.40-7.25 (m,2H), 6.22 (s, 1H), 4.44-4.28 (m, 2H), 3.90 (s, 3H), 3.75-3.70 (m, 1H),3.68-3.60 (m, 1H), 3.45-3.40 (m, 2H), 2.45-2.37 (m, 2H), 1.45-1.31 (m,2H).

Example 21 & Example 22

(4aR,8aR)-1-(5-fluoro-2-methoxy-6-(3-(trifluoromethyl)phenyl)pyridin-3-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(5-fluoro-2-methoxy-6-(3-(trifluoromethyl)phenyl)pyridin-3-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Step 1:

Rac-trans-1-(6-bromo-5-fluoro-2-methoxypyridin-3-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Following the procedure described in Example 1, step 1-4 and makingnon-critical variations as required to replace4-bromo-5-fluoro-2-methoxyaniline with6-bromo-5-fluoro-2-methoxypyridin-3-amine,Rac-trans-1-(6-bromo-5-fluoro-2-methoxypyridin-3-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(290 mg, 45%) was obtained as a yellow solid. LCMS (ESI) m/z: 505.9[M+H]⁺.

Step 2:

Rac-trans-1-(5-fluoro-2-methoxy-6-(3-(trifluoromethyl)phenyl)pyridin-3-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Following the procedure described in Example 1, step 5 and makingnon-critical variations as required to replaceRac-trans-1-(4-bromo-5-fluoro-2-methoxyphenyl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamidewithRac-trans-1-(6-bromo-5-fluoro-2-methoxypyridin-3-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide,(4-fluoro-3-(trifluoromethyl)phenyl)boronic acid with(3-(trifluoromethyl)phenyl)boronic acid,Rac-trans-1-(5-fluoro-2-methoxy-6-(3-(trifluoromethyl)phenyl)pyridin-3-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(150 mg, 55%) as a white solid. LCMS (ESI) m/z: 572.2 [M+H]⁺.

Step 3:

(4aR,8aR)-1-(5-fluoro-2-methoxy-6-(3-(trifluoromethyl)phenyl)pyridin-3-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(5-fluoro-2-methoxy-6-(3-(trifluoromethyl)phenyl)pyridin-3-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Rac-trans-1-(5-fluoro-2-methoxy-6-(3-(trifluoromethyl)phenyl)pyridin-3-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(150 mg) was separated by using chiral SFC (Chiralpak IC (250 mm*30 mm,5 um), Supercritical CO₂/EtOH+0.1% NH₄OH=55/45; 60 mL/min) to give(4aR,8aR)-1-(5-fluoro-2-methoxy-6-(3-(trifluoromethyl)phenyl)pyridin-3-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(55 mg, first peak) as a white solid and(4aS,8aS)-1-(5-fluoro-2-methoxy-6-(3-(trifluoromethyl)phenyl)pyridin-3-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(70 mg, second peak) as a white solid. Absolute configuration wasarbitrarily assigned to each enantiomer. Example 21: LCMS (ESI) m/z:572.2 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.52-8.50 (m, 1H), 8.29 (d,J=7.2 Hz, 1H), 8.25 (s, 1H), 8.09-7.84 (m, 2H), 7.83-7.76 (m, 1H),6.35-6.26 (m, 1H), 4.45-4.27 (m, 2H), 4.02-3.90 (m, 3H), 3.82-3.61 (m,3H), 3.54-3.50 (m, 1H), 2.65-2.55 (m, 2H), 1.47-1.40 (m, 1H), 1.34-1.20(m, 1H). Example 22: LCMS (ESI) m/z: 572.2 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 8.47-8.44 (m, 1H), 8.28 (d, J=7.2 Hz, 1H), 8.25 (s, 1H),8.09-7.84 (m, 2H), 7.83-7.76 (m, 1H), 6.32-6.25 (m, 1H), 4.45-4.27 (m,2H), 4.00-3.91 (m, 3H), 3.82-3.59 (m, 3H), 3.52-3.49 (m, 1H), 2.63-2.54(m, 2H), 1.45-1.39 (m, 1H), 1.32-1.17 (m, 1H).

Example 23 & Example 24

(4aR,8aR)-1-(5-cyano-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(5-cyano-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Step 1:

Rac-trans-tert-butyl4-((5-chloro-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)amino)-3-hydroxypiperidine-1-carboxylate

Following the procedure described in Example 1, step 1 and makingnon-critical variations as required to replace4-bromo-5-fluoro-2-methoxyaniline with5-chloro-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-amine,rac-trans-tert-butyl4-((5-chloro-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)amino)-3-hydroxypiperidine-1-carboxylate(200 mg, 37%) was obtained as yellow oil. ¹H NMR (400 MHz, CDCl₃) δ 7.61(s, 1H), 7.53 (d, J=7.6 Hz, 1H), 7.46-7.36 (m, 2H), 7.23 (d, J=7.6 Hz,1H), 6.55 (d, J=13.2 Hz, 1H), 4.39 (s, 1H), 4.18-4.09 (m, 1H), 3.95-3.85(m, 1H), 3.55-3.43 (m, 1H), 3.31-3.18 (m, 1H), 2.97-2.70 (m, 2H),2.08-1.97 (m, 1H), 1.37 (s, 9H), 1.36-1.34 (m, 1H).

Step 2:

Rac-trans-tert-butyl1-(5-chloro-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-carboxylate

Following the procedure described in Example 1, step 2 and makingnon-critical variations as required to replace rac-trans-tert-butyl4-((4-bromo-5-fluoro-2-methoxyphenyl)amino)-3-hydroxypiperidine-1-carboxylatewith rac-trans-tert-butyl4-((5-chloro-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)amino)-3-hydroxypiperidine-1-carboxylate,rac-trans-tert-butyl1-(5-chloro-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-carboxylate(6.1 g, 85%) was obtained as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ7.78 (s, 1H), 7.75-7.67 (m, 2H), 7.65-7.56 (m, 2H), 7.18-7.05 (m, 1H),4.63-4.35 (m, 3H), 4.31-4.17 (m, 1H), 3.95-3.53 (m, 2H), 2.86-2.54 (m,2H), 1.48 (s, 9H), 1.47-1.45 (m, 2H).

Step 3:

Rac-trans-tert-butyl1-(5-cyano-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-carboxylate

To a solution of Race-trans-tert-butyl1-(5-chloro-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-carboxylate(400 mg, 0.76 mmol), KOAc (223 mg, 2.27 mmol) and K₄Fe(CN)₆ 3H₂O (223mg, 0.38 mmol) in 1,4-dioxane (4 mL) and water (4 mL) were added2-di-tert-butylphosphino-2′,4′,6′-triisopropylbiphenyl (32 mg, 0.08mmol) and[(2-di-tert-butylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′-biphenyl)]palladium(II) methanesulfonate (60 mg, 0.08 mmol). The reaction mixturewas heated to 100° C. for 16 h under a nitrogen atmosphere. Aftercooling to room temperature, EtOAc (30 mL) was added, and washed withbrine (10 mL). The organic layer was dried over anhydrous Na₂SO₄,filtered and concentrated in vacuo. The crude residue was purified bysilica gel chromatography (solvent gradient: 0-25% EtOAc in petroleumether) to give the title compound (200 mg, 51%) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ 7.86 (d, J=7.6 Hz, 1H), 7.79-7.62 (m, 4H),7.24-7.17 (m, 1H), 4.68-4.36 (m, 3H), 4.31-4.01 (m, 1H), 3.83-3.55 (m,2H), 2.90-2.60 (m, 2H), 1.50-1.40 (m, 11H)

Step 4:

Rac-trans-6-fluoro-4-((4aR,8aR)-2-oxooctahydro-1H-pyrido[3,4-b][1,4]oxazin-1-yl)-3′-(trifluoromethyl)-[1,1′-biphenyl]-3-carbonitrile

To a solution of rac-trans-tert-butyl1-(5-cyano-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-carboxylatein EtOAc (4 mL) was added 4M HCl (0.48 mL, 1.92 mmol) in EtOAc. Thesolution was stirred at room temperature for 2 h. The reaction mixturewas quenched with sat. aq. NaHCO₃ (30 mL) and extracted with EtOAc (30mL). The organic layer was dried over anhydrous Na₂SO₄, filtered andconcentrated in vacuo to give the title compound (180 mg, crude) as ayellow solid that required no further purification.

Step 5:

Rac-trans-1-(5-cyano-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Following the procedure described in Example 1, step 4 and makingnon-critical variations as required to replacerac-trans-1-(4-bromo-5-fluoro-2-methoxyphenyl)hexahydro-1H-pyrido[3,4-b][1,4]oxazin-2(3H)-onewithrac-trans-6-fluoro-4-((4aR,8aR)-2-oxooctahydro-1H-pyrido[3,4-b][1,4]oxazin-1-yl)-3′-(trifluoromethyl)-[1,1′-biphenyl]-3-carbonitrile,rac-trans-1-(5-cyano-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(120 mg, 49%) was obtained as yellow oil. LCMS (ESI) m/z: 565.7 [M+H]⁺.

(4aR,8aR)-1-(5-cyano-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(5-cyano-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Rac-trans-1-(5-cyano-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(120 mg) was separated by using chiral SFC (Phenomenex C2 (250 mm*30 mm,5 um), Supercritical CO₂/EtOH+0.1% NH₄OH=60/40; 55 mL/min) to give(4aR,8aR)-1-(5-cyano-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(25 mg, first peak) as a white solid and(4aS,8aS)-1-(5-cyano-2-fluoro-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(23 mg, second peak) as a white solid. Absolute configuration wasarbitrarily assigned to each enantiomer. Example 23: LCMS (ESI) m/z:566.1 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 11.23 (s, 1H), 8.75 (s, 1H),8.42-8.30 (m, 1H), 8.01 (s, 1H), 7.96 (d, J=8.0 Hz, 1H), 7.89-7.64 (m,3H), 6.40 (s, 1H), 4.52-4.40 (m, 2H), 4.08-3.98 (m, 1H), 3.94-3.82 (m,1H), 3.81-3.67 (m, 1H), 3.66-3.58 (m, 1H), 2.88-2.75 (m, 2H), 1.68-1.45(m, 1H), 1.34-1.19 (m, 1H). Example 24: LCMS (ESI) m/z: 566.1 [M+H]⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 11.23 (s, 1H), 8.75 (s, 1H), 8.42-8.30 (m, 1H),8.01 (s, 1H), 7.96 (d, J=8.0 Hz, 1H), 7.89-7.64 (m, 3H), 6.40 (d, J=1.6Hz, 1H), 4.52-4.40 (m, 2H), 4.08-3.98 (m, 1H), 3.94-3.82 (m, 1H),3.81-3.67 (m, 1H), 3.66-3.58 (m, 1H), 2.88-2.75 (m, 2H), 1.68-1.45 (m,1H), 1.34-1.19 (m, 1H).

Example 25 & Example 26

(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl-N-(isoxazol-3-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Step 1:

Rac-trans-tert-butyl1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-carboxylate

To a solution of Rac-trans-tert-butyl1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-carboxylate(1.76 g, 3.36 mmol) in tetrahydrofuran (20 mL) was addedboranedimethylsulfide complex (1.01 mL, 10.07 mmol) at 0° C. under anitrogen atmosphere. Then the mixture was stirred at room temperaturefor 16 h. The mixture was quenched with MeOH (20 mL) dropwise, and thenheated to 80° C. for 1 h. The reaction mixture was concentrated in vacuoand the crude residue was purified by silica gel chromatography (solventgradient: 10-20% EtOAc in petroleum ether) to give the title compound(1.27 g, 74%) as colorless oil.

Step 2:

Rac-trans-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)octahydro-1H-pyrido[3,4-b][1,4]oxazinehydrochloride

To a solution of Rac-trans-tert-butyl1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-carboxylate(600 mg, 1.18 mmol) in EtOAc (5 mL) was added 4M HCl (1.47 mL, 5.88mmol) in EtOAc. The reaction mixture was stirred at room temperature for2 h. The reaction mixture was concentrated in vacuo to give the titlecompound (480 mg, crude) as colorless oil that required no furtherpurification. LCMS (ESI) m/z: 411.2 [M+H]⁺.

Step 3:

Rac-trans-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)hexahydro-H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

To a solution of imidazole (1.66 g, 24.37 mmol) and 3-aminoisoxazole(0.66 g, 7.8 mmol) in DCM (15 ml) was added SO₂Cl₂ (0.64 mL, 7.8 mmol)at −78° C. After stirring at room temperature for 30 min,rac-trans-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)octahydro-1H-pyrido[3,4-b][1,4]oxazinehydrochloride (400 mg, 0.97 mmol) was added. The mixture was heated to80° C. for 1 h under a nitrogen atmosphere. The reaction was dilutedwith water (20 mL) and extracted with dichloromethane (20 mL×3). Thecombined organic layers were washed with brine (20 mL×2), dried overanhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue waspurified by prep-TLC (EtOAc/petroleum ether=1/1) to give the titlecompound (180 mg, 29%) as light yellow oil. LCMS (ESI) m/z: 557.2[M+H]⁺.

(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Rac-trans-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(180 mg, 0.32 mmol) was separated by using chiral SFC (Chiralpak AD (250mm*30 mm, 10 um), Supercritical CO₂/IPA+0.1% NH₄OH=70/30; 50 mL/min) togive(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(32 mg, first peak) as a light yellow solid and(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(45 mg, second peak) as a white solid. Absolute configuration wasarbitrarily assigned to each enantiomer. Example 25: LCMS (ESI) m/z:557.1 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 11.13 (s, 1H), 8.75 (d, J=1.6Hz, 1H), 7.86-7.90 (m, 2H), 7.68-7.77 (m, 2H), 7.17 (d, J=12.0 Hz, 1H),7.12 (d, J=8.0 Hz, 1H), 6.40 (d, J=1.6 Hz, 1H), 3.84-3.85 (m, 4H),3.71-3.78 (m, 1H), 3.64-3.70 (m, 1H), 3.58-3.55 (m, 1H), 3.26-3.31 (m,1H), 3.07 (d, J=8.0 Hz, 1H), 2.87-2.93 (m, 2H), 2.69-2.80 (m, 2H),1.83-1.78 (m, 1H), 1.06-1.13 (m, 1H). Example 26: LCMS (ESI) m/z: 557.1[M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 11.13 (s, 1H), 8.76 (d, J=1.6 Hz,1H), 7.86-7.91 (m, 2H), 7.68-7.78 (m, 2H), 7.18 (d, J=12.0 Hz, 1H), 7.13(d, J=8.0 Hz, 1H), 6.40 (d, J=1.6 Hz, 1H), 3.84-3.85 (s, 4H), 3.71-3.80(m, 1H), 3.64-3.70 (m, 1H), 3.58-3.55 (m, 1H), 3.27-3.31 (m, 1H), 3.07(d, J=12.0 Hz, 1H), 2.88-2.94 (m, 2H), 2.70-2.81 (m, 2H), 1.83-1.78 (m,1H), 1.04-1.16 (m, 1H).

Example 27 & Example 28

(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(pyrimidin-4-yl)hexahydro-H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(pyrimidin-4-yl)hexahydro-H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Step 1:

N-(2,4-dimethoxybenzyl)-N-(pyrimidin-4-yl)-1H-imidazole-1-sulfonamide

To a solution of N-[(2,4-dimethoxyphenyl)methyl]pyrimidin-4-amine (1.0g, 4.08 mmol) and 1,1′-sulfonyldiimidazole (3.23 g, 16.31 mmol) in THF(2 mL) was added lithiumbis(trimethylsilyl)amide (1M, 12.23 mL, 12.23mmol) dropwise. Then the mixture was heated to 80° C. for 3 h under anitrogen atmosphere. The reaction was quenched with water (100 mL),extracted with EtOAc (50 mL×2). The combined organic layers were washedwith brine (100 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified by silica gelchromatography (solvent gradient: 0-5% MeOH in DCM) to give the titlecompound (2.2 g, 10%) as a light yellow solid. LCMS (ESI) m/z: 376.1[M+H]⁺.

Step 2:

Rac-trans-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(pyrimidin-4-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

A 20-mL vial was charged withRac-trans-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)octahydro-1H-pyrido[3,4-b][1,4]oxazine(100 mg, 0.24 mmol),N-(2,4-dimethoxybenzyl)-N-(pyrimidin-4-yl)-1H-imidazole-1-sulfonamide(1.70 g, 0.32 mmol) and N,N-diisopropylethylamine (110 mg, 0.85 mmol) inacetonitrile (10 mL). The vial was sealed with a PTFE lined cap and heatto 130° C. for 5.5 h. After cooling to room temperature, the mixture wasconcentrated in vacuo. The residue was diluted with EtOAc (30 mL). Theorganic layer was washed with sat. aq. citric acid (20 mL), brine (20mL), dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo.The residue was purified by reverse phase chromatography (acetonitrile54-84%/0.225% formic acid in water) to give the title compound (10 mg,7%) as a white solid. LCMS (ESI) m/z: 568.1 [M+H]⁺.

Step 3:

(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(pyrimidin-4-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(pyrimidin-4-yl)hexahydro-H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Rac-trans-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(pyrimidin-4-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(10 mg) was separated by using chiral SFC (Phenomenex C2 (250 mm*30 mm,10 um), Supercritical CO₂/EtOH+0.1% NH₄OH=50/50; 60 mL/min) to give(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(pyrimidin-4-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(5 mg, first peak) as a white solid and(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(pyrimidin-4-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(3 mg, second peak) as a white solid. Absolute configuration wasarbitrarily assigned to each enantiomer. Example 27: LCMS (ESI) m/z:568.1 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.45 (s, 1H), 8.08 (s, 1H),7.92-7.84 (m, 2H), 7.79-7.67 (m, 2H), 7.21-7.09 (m, 2H), 6.82-6.78 (m,1H), 3.85-3.82 (m, 4H), 3.81-3.70 (m, 1H), 3.62-3.60 (m, 1H), 3.49-3.46(m, 2H), 3.09-3.05 (m, 1H), 2.91-2.57 (m, 4H), 1.80-1.77 (m, 1H),1.17-1.06 (m, 1H). Example 28: LCMS (ESI) m/z: 568.1 [M+H]⁺. ¹H NMR (400MHz, DMSO-d₆) δ 8.47 (s, 1H), 8.10 (s, 1H), 7.92-7.83 (m, 2H), 7.78-7.67(m, 2H), 7.20-7.09 (m, 2H), 6.84-6.81 (m, 1H), 3.88-3.81 (m, 4H),3.80-3.70 (m, 1H), 3.62-3.60 (m, 1H), 3.51-3.47 (m, 2H), 3.08-3.05 (m,1H), 2.90-2.70 (m, 3H), 2.67-2.57 (m, 1H), 1.81-1.78 (m, 1H), 1.18-1.06(m, 1H).

Example 29 & Example 30

(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethoxy)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethoxy)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Following the procedure described in Example 3 and making non-criticalvariations as required to 3-(trifluoromethyl)phenylboronic acid with3-(trifluoromethoxy)phenylboronic acid,(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethoxy)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(first peak) and(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(second peak) were obtained. Absolute configuration was arbitrarilyassigned to each enantiomer. Example 29: LCMS (ESI) m/z: 587.0 [M+H]⁺.¹H NMR (400 MHz, DMSO-d₆) δ 11.16 (s, 1H), 8.77 (dd, J=6.0, 1.8 Hz, 1H),7.76-7.62 (m, 2H), 7.60 (s, 1H), 7.50-7.41 (m, 1H), 7.30-7.15 (m, 2H),6.39 (t, J=2.1 Hz, 1H), 4.43-4.28 (m, 2H), 3.90-3.79 (m, 4H), 3.79-3.66(m, 1H), 3.62 (td, J=9.7, 4.4 Hz, 1H), 2.89-2.69 (m, 2H), 1.45 (dd,J=30.8, 8.5 Hz, 2H). Example 30: LCMS (ESI) m/z: 587.0 [M+H]⁺. ¹H NMR(400 MHz, DMSO-d₆) δ 11.16 (s, 1H), 8.75 (dd, J=6.3, 1.8 Hz, 1H),7.70-7.63 (m, 2H), 7.61 (d, J=9.9 Hz, 1H), 7.50-7.41 (m, 1H), 7.30-7.14(m, 2H), 6.39 (t, J=2.0 Hz, 1H), 4.35 (d, J=15.4 Hz, 2H), 3.89-3.79 (m,4H), 3.78-3.54 (m, 2H), 3.42-3.34 (m, 1H), 2.88-2.64 (m, 2H), 1.56-1.19(m, 2H).

Example 31 & Example 32

(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethoxy)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)hexahydro-H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide&(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethoxy)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide

Following the procedure described in Example 25 and making non-criticalvariations as required to replace rac-trans-tert-butyl1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-carboxylatewith rac-trans-tert-butyl1-(2-fluoro-5-methoxy-3′-(trifluoromethoxy)-[1,1′-biphenyl]-4-yl)-2-oxohexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-carboxylate,(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethoxy)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)hexahydro-H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(first peak) and(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethoxy)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)hexahydro-1H-pyrido[3,4-b][1,4]oxazine-6(7H)-sulfonamide(second peak) were obtained. Example 31: LCMS (ESI) m/z: 573.1 [M+H]⁺.¹H NMR (400 MHz, DMSO-d₆) δ 11.12 (s, 1H), 8.76 (d, J=1.8 Hz, 1H),7.66-7.57 (m, 2H), 7.55 (dd, J=2.3, 1.3 Hz, 1H), 7.42-7.36 (m, 1H), 7.16(d, J=12.1 Hz, 1H), 7.10 (d, J=7.5 Hz, 1H), 6.40 (d, J=1.8 Hz, 1H),3.90-3.84 (m, 1H), 3.84 (s, 3H), 3.75 (td, J=11.4, 2.6 Hz, 1H),3.71-3.65 (m, 1H), 3.62-3.53 (m, 1H), 3.30-3.24 (m, 1H), 3.07 (d, J=11.5Hz, 1H), 2.96-2.86 (m, 2H), 2.83-2.69 (m, 2H), 1.81 (d, J=13.1 Hz, 1H),1.17-1.02 (m, 1H). Example 32: LCMS (ESI) m/z: 573.1 [M+H]⁺. ¹H NMR (400MHz, DMSO-d₆) δ 11.12 (s, 1H), 8.76 (d, J=1.8 Hz, 1H), 7.65-7.57 (m,2H), 7.55 (dd, J=2.5, 1.4 Hz, 1H), 7.40 (ddt, J=6.5, 2.6, 1.2 Hz, 1H),7.16 (d, J=12.1 Hz, 1H), 7.10 (d, J=7.5 Hz, 1H), 6.40 (d, J=1.8 Hz, 1H),3.91-3.85 (m, 1H), 3.84 (s, 3H), 3.75 (td, J=11.5, 2.6 Hz, 1H),3.71-3.64 (m, 1H), 3.62-3.53 (m, 1H), 3.07 (d, J=11.5 Hz, 1H), 2.90 (tt,J=9.3, 2.5 Hz, 2H), 2.74 (t, J=11.2 Hz, 2H), 1.81 (dd, J=13.7, 3.5 Hz,1H), 1.09 (qd, J=11.5, 9.9, 3.0 Hz, 1H).

Example 33 & Example 34

(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-2H-pyrido[4,3-d][1,3]oxazine-6(4H)-sulfonamide&(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-2H-pyrido[4,3-d][1,3]oxazine-6(4H)-sulfonamide

Step 1

rac-trans-tert-butyl2-oxohexahydro-2H-pyrido[4,3-d][1,3]oxazine-6(4H)-carboxylate

To a suspension of rac-trans-octahydropyrido[4,3-d][1,3]oxazin-2-onehydrochloride (500 mg, 2.59 mmol) in MeOH (6 mL) was added TEA (656 mg,6.49 mmol). Boc₂O (680 mg, 3.11 mmol) was added. The mixture was stirredat room temperature for 18 h. The mixture was then concentrated. Theresidue was partitioned between DCM and water. The aqueous layer wasextracted with DCM (2×). The combined organics were dried (Na₂SO₄),filtered and concentrated. The crude product was purified by silicaflash chromatography (0-10% MeOH/DCM) to give the title compound (600mg, 90%) as a white solid. LCMS (ESI) m/z 257 [M+H]⁺.

Step 2:

rac-trans-tert-butyl1-[5-fluoro-2-methoxy-4-[3-(trifluoromethyl)phenyl]phenyl]-2-oxo-4,4a,5,7,8,8a-hexahydropyrido[4,3-d][1,3]oxazine-6-carboxylate

The mixture of rac-trans-tert-butyl2-oxo-4,4a,5,7,8,8a-hexahydro-1H-pyrido[4,3-d][1,3]oxazine-6-carboxylate(410 mg, 1.6 mmol),1-fluoro-5-iodo-4-methoxy-2-[3-(trifluoromethyl)phenyl]benzene (792 mg,2.0 mmol), CuI (335 mg, 1.76 mmol), (+/−)-TRANS-1,2-DIAMINOCYCLOHEXANE(183 mg, 1.6 mmol) and CsF (608 mg, 4.0 mmol) in 2-MeTHF (10 mL) washeated at 100° C. for 18 h then at 120° C. for 48. The mixture wasfiltered off celite, concentrated. The crude product was purified bysilica flash chromatography (0-100% iPrOAc/heptane) to give the titlecompound (160 mg, 19%) as brown oil. LCMS (ESI) m/z 525 [M+H]⁺.

Step 3:

rac-trans-1-[5-fluoro-2-methoxy-4-[3-(trifluoromethyl)phenyl]phenyl]-4a,5,6,7,8,8a-hexahydro-4H-pyrido[4,3-d][1,3]oxazin-2-one

To a solution of rac-trans-tert-butyl1-[5-fluoro-2-methoxy-4-[3-(trifluoromethyl)phenyl]phenyl]-2-oxo-4,4a,5,7,8,8a-hexahydropyrido[4,3-d][1,3]oxazine-6-carboxylate(160 mg, 0.301 mmol) in 1,4-dioxane (4 mL) was added HCl (4 M indioxane, 1.9 mL). The mixture was stirred at room temperature for 17 h.The mixture was concentrated, diluted with DCM, washed with sat. NaHCO₃.The aqueous layer was extracted with DCM (2×). The combined organicswere dried (Na₂SO₄), filtered and concentrated to give the titlecompound (130 mg, 100%) as a yellow oil which was used directly in thenext step. LCMS (ESI) m/z 425 [M+H]⁺.

Step 4:

rac-trans-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-2H-pyrido[4,3-d][1,3]oxazine-6(4H)-sulfonamide

To a solution of isoxazol-3-amine (128 mg, 1.53 mmol) and imidazole (311mg, 4.58 mmol) in DCM (4 mL) at −78° C. was added SO₂Cl₂ (206 mg, 1.53mmol) dropwise. The mixture was warmed to room temperature and stirredfor 30 min. A solution ofrac-trans-1-[5-fluoro-2-methoxy-4-[3-(trifluoromethyl)phenyl]phenyl]-4a,5,6,7,8,8a-hexahydro-4H-pyrido[4,3-d][1,3]oxazin-2-one(129 mg, 0.305 mmol) in DCM (1 mL) was added. The mixture was heated at80° C. for 30 min. The mixture was quenched with water, extracted withDCM (3×). The combined organics were dried (Na₂SO₄), filtered andconcentrated. The crude product was purified by silica flashchromatography (0-10% MeOH/DCM) to give the title compound (121 mg, 69%)as yellow oil. LCMS (ESI) m/z 571 [M+H]⁺.

Step 5:

(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-2H-pyrido[4,3-d][1,3]oxazine-6(4H)-sulfonamide&(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-2H-pyrido[4,3-d][1,3]oxazine-6(4H)-sulfonamide

Rac-trans-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-2H-pyrido[4,3-d][1,3]oxazine-6(4H)-sulfonamidewas separated by using chiral SFC (Whelko-01 (150 mm*21.2 mm, 5 um),Supercritical CO₂/MeOH+0.1% NH₄OH=60/40; 80 mL/min) to give(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-2H-pyrido[4,3-d][1,3]oxazine-6(4H)-sulfonamide(first peak) &(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxohexahydro-2H-pyrido[4,3-d][1,3]oxazine-6(4H)-sulfonamide(second peak) as an off-white solid. Absolute configuration wasarbitrarily assigned to each enantiomer. Example 33: LCMS (ESI) m/z:571.0 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 11.08 (s, 1H), 8.75 (dd,J=5.7, 1.7 Hz, 1H), 7.92 (d, J=5.7 Hz, 2H), 7.81 (d, J=7.8 Hz, 1H), 7.75(t, J=7.9 Hz, 1H), 7.47-7.08 (m, 2H), 6.39 (d, J=1.8 Hz, 1H), 4.40-4.29(m, 1H), 4.11 (p, J=10.6 Hz, 1H), 3.94-3.71 (m, 4H), 3.69-3.52 (m, 2H),2.80-2.57 (m, 2H), 2.24-1.95 (m, 1H), 1.48-1.12 (m, 2H). Example 34:LCMS (ESI) m/z: 571.0 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 11.08 (s, 1H),8.73 (dd, J=6.0, 1.8 Hz, 1H), 7.92 (d, J=6.1 Hz, 2H), 7.86-7.68 (m, 2H),7.47-7.05 (m, 2H), 6.38 (d, J=1.8 Hz, 1H), 4.43-4.24 (m, 1H), 4.12 (q,J=11.0 Hz, 1H), 3.85 (d, J=26.2 Hz, 4H), 3.71-3.47 (m, 2H), 2.78-2.53(m, 2H), 2.22-1.95 (m, 1H), 1.49-1.10 (m, 1H).

Examples 35, 36, 37 & 38

(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxooctahydro-1,6-naphthyridine-6(2H)-sulfonamide&(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxooctahydro-1,6-naphthyridine-6(2H)-sulfonamide&(4aS,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxooctahydro-1,6-naphthyridine-6(2H)-sulfonamide&(4aR,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxooctahydro-1,6-naphthyridine-6(2H)-sulfonamide

Step 1:

benzyl 3-(3-ethoxy-3-oxo-propyl)-4-oxo-piperidine-1-carboxylate

The mixture of N-benzyloxycarbonyl-4-piperidone (5 g, 21.4 mmol),pyrrolidine (2.29 g, 32.2 mmol) in benzene (60 mL) was heated at refluxfor 20 h with a Dean-Stark trap. The mixture was then concentrated. Theresidue was again dissolved in benzene (60 mL) and ethyl acrylate (4.29g, 42.9 mmol) was added. The mixture was heated at reflux for 24 h, thencooled to room temperature and filtered off solid. The filtrate wasconcentrated. The crude product was purified by silica flashchromatography with an ELSD detector (0-75% iPrOAc/heptane) to give thetitle compound (5.22 g, 73%) as yellow oil. ¹H NMR (400 MHz,Chloroform-d) δ 7.43-7.30 (m, 5H), 5.23-5.11 (m, 2H), 4.31-4.05 (m, 3H),3.47-3.35 (m, 1H), 3.07 (s, 1H), 2.59-2.28 (m, 5H), 2.14-2.02 (m, 1H),1.66-1.58 (m, 2H), 1.32-1.17 (m, 3H).

Step 2:

3-(1-benzyloxycarbonyl-4-oxo-3-piperidyl)propanoic Acid

The mixture of benzyl3-(3-ethoxy-3-oxo-propyl)-4-oxo-piperidine-1-carboxylate (1.0 g, 3.0mmol) in THF (35 mL), MeOH (3 mL) and LiOH (3 M, 4 mL) was stirred atroom temperature for 3 h. The volatiles were removed under reducedpressure. The aqueous residue was acidified with 2 N HCl to pH ˜3,extracted with iPrOAc. The combined organics were dried (Na₂SO₄),filtered and concentrated to give the title compound (950 mg, quant.yield) as yellow oil. ¹H NMR (400 MHz, DMSO-d₆) δ 12.08 (s, 1H),7.49-7.23 (m, 5H), 5.13 (s, 2H), 4.14-3.84 (m, 2H), 3.50-3.36 (m, 1H),3.23-2.95 (m, 1H), 2.48-2.42 (m, 2H), 2.39-2.29 (m, 1H), 2.24 (t, J=7.6Hz, 2H), 1.85 (dq, J=14.4, 7.3 Hz, 1H), 1.56-1.29 (m, 1H).

3-[1-benzyloxycarbonyl-4-(4-bromo-5-fluoro-2-methoxy-anilino)-3-piperidyl]propanoicAcid

To a screw-capped vial was added3-(1-benzyloxycarbonyl-4-oxo-3-piperidyl)propanoic acid (810 mg, 2.653mmol), 4-bromo-5-fluoro-2-methoxy-aniline (584 mg, 2.653 mmol), toluene(5.306 mL) and phenylsilane (574 mg, 5.306 mmol). The mixture was heatedat 100° C. for 3 h. The mixture was diluted with iPrOAc, washed with 2 MNa₂CO₃. The combined organics were dried (Na₂SO₄), filtered andconcentrated. The crude product was purified by silica flashchromatography (0-10% MeOH/DCM) to give the title compound (1.137 g,84%) as brown oil. LCMS (ESI) m/z 509 [M+H]⁺.

Step 4:

benzyl1-(4-bromo-5-fluoro-2-methoxy-phenyl)-2-oxo-4,4a,5,7,8,8a-hexahydro-3H-1,6-naphthyridine-6-carboxylate

3-[1-benzyloxycarbonyl-4-(4-bromo-5-fluoro-2-methoxy-anilino)-3-piperidyl]propanoicacid (1.137 g, 2.232 mmol) was dissolved in DMF (6 mL). HATU (934 mg,2.455 mmol) was added followed by DIPEA (432.7 mg, 3.348 mmol). Themixture was stirred at room temperature for 4 h. The reaction wasquenched with 2 M Na₂CO₃, extracted with iPrOAc (2×). The combinedorganics were dried (Na₂SO₄), filtered and concentrated. The crudeproduct was purified by silica flash chromatography (0-100%iPrOAc/heptane) to give the title compound (920 mg, 84%) as brown foam.LCMS (ESI) m/z 491 [M+H]⁺. ¹H NMR (400 MHz, Chloroform-d) δ 7.44-7.29(m, 5H), 7.10 (dd, J=8.3, 6.1 Hz, 1H), 6.89 (dd, J=29.0, 8.2 Hz, 1H),5.24-5.05 (m, 2H), 4.26-4.06 (m, 2H), 3.87-3.72 (m, 3H), 3.69-3.54 (m,1H), 3.20-3.04 (m, 1H), 2.76-2.45 (m, 3H), 2.40-2.20 (m, 1H), 2.12-2.02(m, 1H), 1.90-1.65 (m, 3H).

Step 5:

benzyl1-[5-fluoro-2-methoxy-4-[3-(trifluoromethyl)phenyl]phenyl]-2-oxo-4,4a,5,7,8,8a-hexahydro-3H-1,6-naphthyridine-6-carboxylate

The mixture of 3-(trifluoromethyl)phenylboronic acid (480 mg, 2.527mmol), benzyl1-(4-bromo-5-fluoro-2-methoxy-phenyl)-2-oxo-4,4a,5,7,8,8a-hexahydro-3H-1,6-naphthyridine-6-carboxylate(920 mg, 1.872 mmol), Pd (PPh₃)₄(216 mg, 0.1872 mmol) and K₂CO₃ (776 mg,5.617 mmol) in 1,4-dioxane (9 mL) and water (3 mL) was heated at 100° C.under N₂ for 1 h. The reaction mixture was diluted with water, extractedwith iPrOAc (3×). The combined organics were dried (Na₂SO₄), filteredand concentrated. The crude product was purified by silica flashchromatography (0-100% iPrOAc/heptane,) to give the title compound (1.1g, 106%) as off-white foam. LCMS (ESI) m/z 557 [M+H]⁺.

Step 6:

1-[5-fluoro-2-methoxy-4-[3-(trifluoromethyl)phenyl]phenyl]-3,4,4a,5,6,7,8,8a-octahydro-1,6-naphthyridin-2-one

The mixture solution of benzyl1-[5-fluoro-2-methoxy-4-[3-(trifluoromethyl)phenyl]phenyl]-2-oxo-4,4a,5,7,8,8a-hexahydro-3H-1,6-naphthyridine-6-carboxylate(560 mg, 1.006 mmol) and Pd(OH)₂ on carbon (494.5 mg, 0.3521 mmol) inEtOAc (5 mL) was stirred at room temperature under H₂ balloon for 14 h.The mixture was filtered off solid, washed with EtOAc, concentrated. Thecrude product was purified by silica flash chromatography (0-10%MeOH/DCM with 1% NH₄OH) to give the title compound (315 mg, 74%) asbrown oil. LCMS (ESI) m/z 423 [M+H]⁺.

Step 7:

1-[5-fluoro-2-methoxy-4-[3-(trifluoromethyl)phenyl]phenyl]-N-isoxazol-3-yl-2-oxo-4,4a,5,7,8,8a-hexahydro-3H-1,6-naphthyridine-6-sulfonamide

To a solution of isoxazol-3-amine (313 mg, 3.729 mmol) and imidazole(761 mg, 11.19 mmol) in DCM (9 mL) at −78° C. was added SO₂Cl₂ (503 mg,3.729 mmol) dropwise. The mixture was warmed to room temperature andstirred for 30 min.1-[5-fluoro-2-methoxy-4-[3-(trifluoromethyl)phenyl]phenyl]-3,4,4a,5,6,7,8,8a-octahydro-1,6-naphthyridin-2-one(315 mg, 0.7457 mmol) in DCM (1 mL) was then added. The mixture washeated at 80° C. for 30 min. The reaction was quenched with water,extracted with DCM (3×). The combined organics were dried (Na₂SO₄),filtered and concentrated. The crude product was purified by silicaflash chromatography (0-10% MeOH/DCM) to give the title compound (235mg, 55%) as yellow oil. LCMS (ESI) m/z 569 [M+H]⁺.

Step 8:

(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxooctahydro-1,6-naphthyridine-6(2H)-sulfonamide&(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxooctahydro-1,6-naphthyridine-6(2H)-sulfonamide&(4aS,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxooctahydro-1,6-naphthyridine-6(2H)-sulfonamide&(4aR,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxooctahydro-1,6-naphthyridine-6(2H)-sulfonamide

1-[5-fluoro-2-methoxy-4-[3-(trifluoromethyl)phenyl]phenyl]-N-isoxazol-3-yl-2-oxo-4,4a,5,7,8,8a-hexahydro-3H-1,6-naphthyridine-6-sulfonamidewas separated by using chiral SFC (Chiralpak ID (150 mm*21.2 mm, 5 um),Supercritical CO₂/MeOH+0.1% NH₄OH=70/30, 70 mL/min) to give(4aR,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxooctahydro-1,6-naphthyridine-6(2H)-sulfonamide(first peak) &(4aS,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxooctahydro-1,6-naphthyridine-6(2H)-sulfonamide(second peak) &(4aS,8aR)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxooctahydro-1,6-naphthyridine-6(2H)-sulfonamide(third peak) &(4aR,8aS)-1-(2-fluoro-5-methoxy-3′-(trifluoromethyl)-[1,1′-biphenyl]-4-yl)-N-(isoxazol-3-yl)-2-oxooctahydro-1,6-naphthyridine-6(2H)-sulfonamide(fourth peak) as off-white solids. Absolute configuration wasarbitrarily assigned to each isomer. Example 35: LCMS (ESI) m/z: 569.1[M+H]. ¹H NMR (400 MHz, DMSO-d₆) δ 11.04 (s, 1H), 8.74 (dd, J=3.4, 1.8Hz, 1H), 7.98-7.87 (m, 2H), 7.85-7.66 (m, 2H), 7.35-7.00 (m, 2H), 6.39(d, J=1.8 Hz, 1H), 3.81 (d, J=22.3 Hz, 3H), 3.72 (t, J=9.6 Hz, 1H), 3.61(t, J=12.7 Hz, 1H), 3.53-3.36 (m, 1H), 3.29-3.17 (m, 1H), 2.78-2.57 (m,2H), 2.47-2.38 (m, 1H), 1.88-1.71 (m, 2H), 1.68-1.14 (m, 3H). Example36: LCMS (ESI) m/z: 569.1 [M+H]. ¹H NMR (400 MHz, DMSO-d₆) δ 11.06 (s,1H), 8.75 (d, J=1.8 Hz, 1H), 7.92 (d, J=6.5 Hz, 2H), 7.86-7.66 (m, 3H),7.24 (d, J=7.1 Hz, 1H), 6.39 (d, J=1.8 Hz, 1H), 3.82 (s, 3H), 3.62-3.47(m, 2H), 3.14-2.97 (m, 1H), 2.74-2.60 (m, 1H), 2.47-2.25 (m, 2H),2.12-1.90 (m, 1H), 1.90-1.51 (m, 3H). Example 37: LCMS (ESI) m/z: 569.1[M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 11.05 (s, 1H), 8.66 (d, J=5.6 Hz,1H), 7.92 (d, J=6.8 Hz, 2H), 7.86-7.66 (m, 2H), 7.35-7.00 (m, 2H), 6.37(d, J=1.8 Hz, 1H), 3.81 (d, J=22.7 Hz, 3H), 3.76-3.52 (m, 2H), 3.51-3.39(m, 2H), 2.72-2.53 (m, 2H), 2.46-2.36 (m, 1H), 1.87-1.69 (m, 2H),1.69-1.12 (m, 3H). Example 38: LCMS (ESI) m/z: 569.1 [M+H]⁺. ¹H NMR (500MHz, DMSO-d₆) δ 10.86 (s, 1H), 8.69 (d, J=1.8 Hz, 1H), 7.90 (d, J=9.0Hz, 2H), 7.78 (d, J=7.9 Hz, 1H), 7.73 (t, J=7.6 Hz, 1H), 7.23 (d, J=7.2Hz, 1H), 7.18 (d, J=10.5 Hz, 1H), 6.37 (d, J=1.7 Hz, 1H), 3.82 (s, 3H),3.76-3.63 (m, 1H), 3.59-3.50 (m, 2H), 3.08 (dd, J=12.5, 3.3 Hz, 1H),2.69 (td, J=12.1, 3.0 Hz, 1H), 2.46-2.38 (m, 2H), 2.38-2.33 (m, 1H),2.04 (td, J=13.3, 12.1, 6.3 Hz, 1H), 1.80-1.73 (m, 1H), 1.71-1.60 (m,2H).

Although the preparation of the free-base or a specific salt form may beillustrated in the Examples above, it is understood that the free-baseand its acid or base salt forms can be interconverted using standardtechniques

Example 39A. Tritiated Compound Binding to Membranes Isolated from Cellsthat Heterologously Express hNav1.7 and the β1 Subunit

Preparation of membranes containing recombinantly expressed sodiumchannels: Frozen recombinant cell pellets were thawed on ice and dilutedto 4 times the cell pellet weight with ice cold 50 mM Tris HCl, pH 7.4buffer. The cell suspensions were homogenized on ice using a motorizedglass dounce homogeniser. Homogenates were further diluted 8.4 timeswith ice cold 50 mM Tris HCl, pH 7.4 buffer and then centrifuged at200×g at 4° C. for 15 min. The supernatants were collected andcentrifuged at 10000×g at 4° C. for 50 min. The pellets were thenre-suspended in 100 mM NaCl, 20 mM Tris HCl, pH 7.4 buffer containing 1%v/v protease inhibitors (Calbiochem) and re-homogenized on ice. Thehomogenized membranes were then processed through a syringe equippedwith a 26 gauge needle. Protein concentrations were determined byBradford Assay and the membranes were stored at −80° C.

Radioligand Binding Studies:

Saturation experiments. A competitive NaV1.7 inhibitor having a methylgroup was tritiated. Three tritiums were incorporated in place of methylhydrogens to generate [³H]compound. Binding of this radioligand wasperformed in 5 mL borosilicate glass test tubes at room temperature.Binding was initiated by adding membranes to increasing concentrationsof [³H]compound in 100 mM NaCl, 20 mM Tris HCl, pH 7.4 buffer containing0.01% w/v bovine serum albumin (BSA) for 18 h. Non-specific binding wasdetermined in the presence of 1 μM unlabeled compound. After 18 h, thereactants were filtered through GF/C glass fiber filters presoaked in0.5% w/v polyethylene imine. Filters were washed with 15 mL ice cold 100mM NaCl, 20 mM Tris HCl, pH7.4 buffer containing 0.25% BSA to separatebound from free ligand. [³H]compound bound to filters was quantified byliquid scintillation counting.

Competitive Binding Experiments:

Binding reactions were performed in 96-well polypropylene plates at roomtemperature for 18 h. In 360 μL, membranes were incubated with 100 pM[³H]compound and increasing concentrations of Test Compound.Non-specific binding was defined in the presence of 1 μM unlabeledcompound. Reactions were transferred and filtered through 96-well glassfiber/C filter plates presoaked with 0.5% polyethylene imine. Thefiltered reactions were washed 5 times with 200 μL ice cold buffercontaining 0.25% BSA. Bound radioactivity was determined by liquidscintillation counting.

Data Analysis:

For saturation experiments, non-specific binding was subtracted fromtotal binding to provide specific binding and these values wererecalculated in terms of pmol ligand bound per mg protein. Saturationcurves were constructed and dissociation constants were calculated usingthe single site ligand binding model: Beq=(Bmax*X)/(X+Kd), where Beq isthe amount of ligand bound at equilibrium, Bmax is the maximum receptordensity, Kd is the dissociation constant for the ligand, and X is thefree ligand concentration. For competition studies percent inhibitionwas determined and IC₅₀ values were calculated using a 4 parameterlogistic model (% inhibition=(A+((B−A)/(1+((x/C){circumflex over( )}D)))) using XLfit, where A and B are the maximal and minimuminhibition respectively, C is the IC₅₀ concentration and D is the (Hill)slope.

Example 39B. Tritiated Compound Binding to Membranes Isolated from Cellsthat Heterologous Express hNav1.7 and the β1 Subunit

Preparation of Membranes Containing Recombinantly Expressed SodiumChannels:

Following the procedure described in Example 39A, and makingmodifications as required to: centrifuge supernatants at 100000×ginstead of 10000×g; re-suspend pellets in a buffer containing 0.01% BSAin addition to 100 mM NaCl, 20 mM Tris HCl, pH 7.4 buffer, and 1% v/vproteasome inhibitors; and storing membranes at −80° C. in single usealiquots instead of pooled.

Radioligand Binding Studies:

Saturation experiments: Following the procedure described in Example243A, and making modifications as required to incubate [3H]compound for3 hours instead of 18 hours.

Competitive Binding Experiments:

Following the procedure described in Example 243A, and makingmodifications as required to: perform binding experiments at roomtemperature for 3 hours instead of 18 hours; use 240 uL of solution and300 pM of [3H]compound instead of 360 uL and 100 pM of [3H]compound; andwash filtered reactions 2 times instead of 5 times.

Data Analysis:

Data was analysed following the procedure described in Example 243A.

Example 39C. Electrophysiological Assay (EP) (In Vitro Assay)

Patch voltage clamp electrophysiology allows for the direct measurementand quantification of block of voltage-gated sodium channels (NaV's),and allows the determination of the time- and voltage-dependence ofblock which has been interpreted as differential binding to the resting,open, and inactivated states of the sodium channel (Hille, B., Journalof General Physiology (1977), 69: 497-515).

The following voltage clamp electrophysiology studies were performed onrepresentative compounds using cells heterologously expressing Nav1.7 orNav1.5 channels. cDNAs for Nav1.7 (NM_002977) and Nav1.5 (AC137587) werestably expressed in Chinese Hamstr Ovary (CHO) cells and CHL (ChineseHamster Lung) cells respectively. Sodium currents were measured in thewhole-cell configuration using Syncropatch 384PE (Nanlon Technologies,Germany). 1NPC®-384 chips with custom medium resistance and single holemode are used. Internal solution consists of (in mM): 110 CsCl, 10 CsCl,20 EGTA, and 10 Hepes (pH adjusted to 7.2); and external solutioncontains (in mM): 60 NMDG, 80 NaCl, 4 KCl, 1 MgCl₂, 2 CaCl₂, 2 D-Glucosemonohydrate, 10 Hepes (pH adjusted to 7.4 with NaOH).

After system flushing, testing compounds are dissolved in externalsolution containing 0.1% Pluronic F-127. The chip is moved into themeasuring head and the instrument primes the chip with external andinternal solutions. 10 l cells are added to the chip from a cell hotel,and a negative pressure of −50 mBar is applied to form a seal. Followingtreatment with seal enhancer solution and wash-off with externalsolution, negative pressure of −250 mbar is applied for 1 second toachieve the whole-cell configuration, followed by three washing steps inexternal solution. 20 l of compounds is added to 40 l in each well (1:3dilution of compounds), and after mixing, 20 l is removed so the volumeis retained at 40 ul. After approximately 13 minutes recordings, 20l/well of 2 uM TTX, or 333 uM Tetracaine (for Nav1.5) is added toachieve full block.

For voltage protocol, an holding potential of −50 mV is applied duringthe whole experiment. A depolarizing step is applied to −10 mV for 10ms, followed by a hyperpolarization step to −150 mV for 20 ms to allowchannel recovery from inactivation. A second depolarizing step isapplied from −150 mV to −10 mV for 10 ms, where currents were measuredto derive blocking effects of compounds. Inhibition is determined basedon 7.5 min of compound incubation.

Data for representative compounds is provided in Table 1.

TABLE 1 Nav1.7 EP_SP LBA EVO hNav1.7 EVO Example Structure IC₅₀ (uM)IC50 (uM)  1

0.0443 0.00112  2

0.0384 0.000915  3

0.067 0.00226  4

0.06 0.00352  5

0.079 0.00279  6

0.0529 0.00184  7

0.167 0.00576  8

0.0828 0.00492  9

2.0 0.0194 10

0.716 0.0478 11

0.23 0.0112 12

0.131 0.00683 13

0.471 0.0117 14

0.332 0.00721 15

2.0 0.0467 16

1.39 0.0495 17

0.214 0.00603 18

0.346 0.00586 19

0.831 0.05 20

2.0 0.0398 21

2.0 0.0475 22

0.907 0.0668 23

2.0 0.0501 24

2.0 0.237 25

0.208 0.00236 26

0.0636 0.0106 27

0.0212 0.00202 28

0.0263 0.0021 29

0.0623 0.00221 30

0.0356 0.00235 31

0.128 0.00598 32

0.148 0.0109 33

0.112 0.0104 34

2.0 0.196 35

0.0493 0.00288 36

0.0207 0.00219 37

0.761 0.0129 38

0.641 0.0107

Example 40 Analgesia Induced by Sodium Channel Blockers

Heat Induced Tail Flick Latency Test

In this test, the analgesia effect produced by administering a compoundof the invention can be observed through heat-induced tail-flick inmice. The test includes a heat source consisting of a projector lampwith a light beam focused and directed to a point on the tail of a mousebeing tested. The tail-flick latencies, which are assessed prior to drugtreatment, and in response to a noxious heat stimulus, i.e., theresponse time from applying radiant heat on the dorsal surface of thetail to the occurrence of tail flick, are measured and recorded at 40,80, 120, and 160 minutes.

For the first part of this study, 65 animals undergo assessment ofbaseline tail flick latency once a day over two consecutive days. Theseanimals are then randomly assigned to one of the 11 different treatmentgroups including a vehicle control, a morphine control, and 9 compoundsat 30 mg/Kg are administered intramuscularly. Following doseadministration, the animals are closely monitored for signs of toxicityincluding tremor or seizure, hyperactivity, shallow, rapid or depressedbreathing and failure to groom. The optimal incubation time for eachcompound is determined via regression analysis. The analgesic activityof the test compounds is expressed as a percentage of the maximumpossible effect (% MPE) and is calculated using the following formula:

$\%\mspace{14mu}{MPE}\mspace{14mu}\frac{{{Postdrug}\mspace{14mu}{latency}} - {{Predrug}\mspace{14mu}{latency}}}{{{Cut}\text{-}{off}\mspace{14mu}{time}\mspace{14mu}\left( {10\mspace{20mu} s} \right)} - {{Predrug}\mspace{14mu}{latency}}} \times 100\%$where:

Postdrug latency=the latency time for each individual animal takenbefore the tail is removed (flicked) from the heat source afterreceiving drug.

Predrug latency=the latency time for each individual animal taken beforethe tail is flicked from the heat source prior to receiving drug.

Cut-off time (10 s)=is the maximum exposure to the heat source.

Acute Pain (Formalin Test)

The formalin test is used as an animal model of acute pain. In theformalin test, animals are briefly habituated to the plexiglass testchamber on the day prior to experimental day for 20 minutes. On the testday, animals are randomly injected with the test articles. At 30 minutesafter drug administration, 50 μL of 10% formalin is injectedsubcutaneously into the plantar surface of the left hind paw of therats. Video data acquisition begins immediately after formalinadministration, for duration of 90 minutes.

The images are captured using the Actimetrix Limelight software whichstores files under the *.llii extension, and then converts it into theMPEG-4 coding. The videos are then analyzed using behaviour analysissoftware “The Observer 5.1”, (Version 5.0, Noldus InformationTechnology, Wageningen, The Netherlands). The video analysis isconducted by watching the animal behaviour and scoring each according totype, and defining the length of the behaviour (Dubuisson and Dennis,1977). Scored behaviours include: (1) normal behaviour, (2) putting noweight on the paw, (3) raising the paw, (4) licking/biting or scratchingthe paw. Elevation, favoring, or excessive licking, biting andscratching of the injected paw indicate a pain response. Analgesicresponse or protection from compounds is indicated if both paws areresting on the floor with no obvious favoring, excessive licking, bitingor scratching of the injected paw.

Analysis of the formalin test data is done according to two factors: (1)Percent Maximal Potential Inhibitory Effect (% MPIE) and (2) pain score.The % MPIEs is calculated by a series of steps, where the first is tosum the length of non-normal behaviours (behaviours 1,2,3) of eachanimal. A single value for the vehicle group is obtained by averagingall scores within the vehicle treatment group. The following calculationyields the MPIE value for each animal:MPIE (%)=100−[(treatment sum/average vehicle value)×100%]

The pain score is calculated from a weighted scale as described above.The duration of the behaviour is multiplied by the weight (rating of theseverity of the response), and divided by the total length ofobservation to determine a pain rating for each animal. The calculationis represented by the following formula:Pain rating=[0(To)+1(T1)+2(T2)+3(T3)]/(To+T1+T2+T3)

CFA Induced Chronic Inflammatory Pain

In this test, tactile allodynia is assessed with calibrated von Freyfilaments. Following a full week of acclimatization to the vivariumfacility, 150 μL of the “Complete Freund's Adjuvant” (CFA) emulsion (CFAsuspended in an oil/saline (1:1) emulsion at a concentration of 0.5mg/mL) is injected subcutaneously into the plantar surface of the lefthind paw of rats under light isoflurane anaesthesia. Animals are allowedto recover from the anaesthesia and the baseline thermal and mechanicalnociceptive thresholds of all animals are assessed one week after theadministration of CFA. All animals are habituated to the experimentalequipment for 20 minutes on the day prior to the start of theexperiment. The test and control articles are administrated to theanimals, and the nociceptive thresholds measured at defined time pointsafter drug administration to determine the analgesic responses to eachof the six available treatments. The time points used are previouslydetermined to show the highest analgesic effect for each test compound.

Thermal nociceptive thresholds of the animals are assessed using theHargreaves test. Animals are placed in a Plexiglas enclosure set on topof an elevated glass platform with heating units. The glass platform isthermostatically controlled at a temperature of approximately 30° C. forall test trials. Animals are allowed to accommodate for 20 minutesfollowing placement into the enclosure until all exploration behaviourceases. The Model 226 Plantar/Tail Stimulator Analgesia Meter (IITC,Woodland Hills, Calif.) is used to apply a radiant heat beam fromunderneath the glass platform to the plantar surface of the hind paws.During all test trials, the idle intensity and active intensity of theheat source are set at 1 and 45 respectively, and a cut off time of 20seconds is employed to prevent tissue damage.

The response thresholds of animals to tactile stimuli are measured usingthe Model 2290 Electrovonfrey anesthesiometer (IITC Life Science,Woodland Hills, Calif.) following the Hargreaves test. Animals areplaced in an elevated Plexiglas enclosure set on a mire mesh surface.After 10 minutes of accommodation, pre-calibrated Von Frey hairs areapplied perpendicularly to the plantar surface of both paws of theanimals in an ascending order starting from the 0.1 g hair, withsufficient force to cause slight buckling of the hair against the paw.Testing continues until the hair with the lowest force to induce a rapidflicking of the paw is determined or when the cut off force ofapproximately 20 g is reached. This cut off force is used because itrepresent approximately 10% of the animals' body weight and it serves toprevent raising of the entire limb due to the use of stiffer hairs,which would change the nature of the stimulus.

Postoperative Models of Nociception

In this model, the hypealgesia caused by an intra-planar incision in thepaw is measured by applying increased tactile stimuli to the paw untilthe animal withdraws its paw from the applied stimuli. While animals areanaesthetized under 3.5% isofluorane, which is delivered via a nosecone, a 1 cm longitudinal incision is made using a number 10 scalpelblade in the plantar aspect of the left hind paw through the skin andfascia, starting 0.5 cm from the proximal edge of the heel and extendingtowards the toes. Following the incision, the skin is apposed using 2,3-0 sterilized silk sutures. The injured site is covered with Polysporinand Betadine. Animals are returned to their home cage for overnightrecovery.

The withdrawal thresholds of animals to tactile stimuli for bothoperated (ipsilateral) and unoperated (contralateral) paws can bemeasured using the Model 2290 Electrovonfrey anesthesiometer (IITC LifeScience, Woodland Hills, Calif.). Animals are placed in an elevatedPlexiglas enclosure set on a mire mesh surface. After at least 10minutes of acclimatization, pre-calibrated Von Frey hairs are appliedperpendicularly to the plantar surface of both paws of the animals in anascending order starting from the 10 g hair, with sufficient force tocause slight buckling of the hair against the paw. Testing continuesuntil the hair with the lowest force to induce a rapid flicking of thepaw is determined or when the cut off force of approximately 20 g isreached. This cut off force is used because it represent approximately10% of the animals' body weight and it serves to prevent raising of theentire limb due to the use of stiffer hairs, which would change thenature of the stimulus.

Neuropathic Pain Model; Chronic Constriction Injury

Briefly, an approximately 3 cm incision is made through the skin and thefascia at the mid thigh level of the animals' left hind leg using a no.10 scalpel blade. The left sciatic nerve is exposed via blunt dissectionthrough the biceps femoris with care to minimize haemorrhagia. Fourloose ligatures are tied along the sciatic nerve using 4-0non-degradable sterilized silk sutures at intervals of 1 to 2 mm apart.The tension of the loose ligatures is tight enough to induce slightconstriction of the sciatic nerve when viewed under a dissectionmicroscope at a magnification of 4 fold. In the sham-operated animal,the left sciatic nerve is exposed without further manipulation.Antibacterial ointment is applied directly into the wound, and themuscle is closed using sterilized sutures. Betadine is applied onto themuscle and its surroundings, followed by skin closure with surgicalclips.

The response thresholds of animals to tactile stimuli are measured usingthe Model 2290 Electrovonfrey anesthesiometer (IITC Life Science,Woodland Hills, Calif.). Animals are placed in an elevated Plexiglasenclosure set on a mire mesh surface. After 10 minutes of accommodation,pre-calibrated Von Frey hairs are applied perpendicularly to the plantarsurface of both paws of the animals in an ascending order starting fromthe 0.1 g hair, with sufficient force to cause slight buckling of thehair against the paw. Testing continues until the hair with the lowestforce to induce a rapid flicking of the paw is determined or when thecut off force of approximately 20 g is reached. This cut off force isused because it represents approximately 10% of the animals' body weightand it serves to prevent raising of the entire limb due to the use ofstiffer hairs, which would change the nature of the stimulus.

Thermal nociceptive thresholds of the animals are assessed using theHargreaves test. Following the measurement of tactile thresholds,animals are placed in a Plexiglass enclosure set on top of an elevatedglass platform with heating units. The glass platform isthermostatically controlled at a temperature of approximately 24 to 26°C. for all test trials. Animals are allowed to accommodate for 10minutes following placement into the enclosure until all explorationbehaviour ceases. The Model 226 Plantar/Tail Stimulator Analgesia Meter(IITC, Woodland Hills, Calif.) is used to apply a radiant heat beam fromunderneath the glass platform to the plantar surface of the hind paws.During all test trials, the idle intensity and active intensity of theheat source are set at 1 and 55 respectively, and a cut off time of 20seconds is used to prevent tissue damage.

Neuropathic Pain Model: Spinal Nerve Ligation

The spinal nerve ligation (SNL) neuropathic pain model is used as ananimal (i.e. rat) model of neuropathic pain. In the SNL test, the lumbarroots of spinal nerves L5 and L6 are tightly ligated to cause nerveinjury, which results in the development of mechanical hyperalgesia,mechanical allodynia and thermal hypersensitivity. The surgery isperformed two weeks before the test day in order for the pain state tofully develop in the animals. Several spinal nerve ligation variationsare used to characterize the analgesic properties of a compound of theinvention.

Ligation of the L5 spinal nerve;

Ligation of the L5 and L6 spinal nerves;

Ligation and transection of the L5 spinal nerve;

Ligation and transection of the L5 and L6 spinal nerves; or

Mild irritation of the L4 spinal nerve in combination with any one ofthe above (1)-(4).

While the animals are anaesthetized under 3.5% isofluorane delivered viaa nose cone, an approximately 2.5 cm longitudinal incision is made usinga number 10 scalpel blade in the skin just lateral to the dorsalmidline, using the level of the posterior iliac crests as the midpointof the incision. Following the incision, the isoflourane is readjustedto maintenance levels (1.5%-2.5%). At mid-sacral region, an incision ismade with the scalpel blade, sliding the blade along the side of thevertebral column (in the saggital plane) until the blade hits thesacrum. Scissors tips are introduced through the incision and the muscleand ligaments are removed from the spine to expose 2-3 cm of thevertebral column. The muscle and fascia are cleared from the spinalvertebra in order to locate the point where the nerve exits from thevertebra. A small glass hook is placed medial to the spinal nerves andthe spinal nerves are gently elevated from the surrounding tissues. Oncethe spinal nerves have been isolated, a small length of non-degradable6-0 sterilized silk thread is wound twice around the ball at the tip ofthe glass hook and passed back under the nerve. The spinal nerves arethen firmly ligated by tying a knot, ensuring that the nerve bulges onboth sides of the ligature. The procedure may be repeated as needed. Insome animals, the L4 spinal nerve may be lightly rubbed (up to 20 times)with the small glass hook to maximize the development of neuropathicpain. Antibacterial ointment is applied directly into the incision, andthe muscle is closed using sterilized sutures. Betadine is applied ontothe muscle and its surroundings, followed by skin closure with surgicalstaples or sterile non-absorable monofilament 5-0 nylon sutures.

The analgesic effect produced by topical administration of a compound ofthe invention to the animals can then be observed by measuring the pawwithdrawal threshold of animals to mechanical tactile stimuli. These maybe measured using either the mechanical allodynia procedure or themechanical hyperalgesia procedure as described below. Afterestablishment of the appropriate baseline measurements by either method,topical formulation of a compound of the invention is applied on theipsilateral ankle and foot. The animals are then placed in plastictunnels for 15 minutes to prevent them from licking the treated area andremoving the compound. Animals are placed in the acrylic enclosure for15 minutes before testing the ipsilateral paw by either of the methodsdescribed below, and the responses are recorded at 0.5, 1.0 and 2.0 hourpost treatment.

Mechanical Allodynia Method

The pain threshold of animals to mechanical alloydnia for both operatedand control animals can be measured approximately 14 days post-surgeryusing manual calibrated von Frey filaments as follows. Animals areplaced in an elevated plexiglass enclosure set on a mire mesh surface.Animals are allowed to acclimate for 20-30 minutes. Pre-calibrated VonFrey hairs are applied perpendicularly to the plantar surface of theipsilateral paw of the animals starting from the 2.0 g hair, withsufficient force to cause slight buckling of the hair against the paw toestablish the baseline measurements. Stimuli are presented in aconsecutive manner, either in an ascending or descending order until thefirst change in response is noted, after which four additional responsesare recorded for a total of six responses. The six responses measured ingrams are entered into a formula as described by Chaplan, S. R. et al.,J. Neurosci. Methods, 1994 July; 53(1):55-63, and a 50% withdrawalthreshold is calculated. This constitutes the mechanical allodyniavalue.

Mechanical Hyperalgesia Method

The response thresholds of animals to tactile stimuli are measured usingthe Model 2290 Electrovonfrey anesthesiometer (IITC Life Science,Woodland Hills, Calif.). Animals are placed in an elevated Plexiglasenclosure set on a wire mesh surface. After 15 minutes of accommodationin this enclosure, a von Frey hair is applied perpendicularly to theplantar surface of the ipsilateral hind paws of the animals, withsufficient force, measured in grams, to elicit a crisp response of thepaw. A response indicates a withdrawal from the painful stimulus andconstitutes the efficacy endpoint. The data are expressed as percentchange from baseline threshold measured in grams.

Example 41 In Vivo Assay for Treatment of Pruritus

The compounds of the invention can be evaluated for their activity asantipruritic agents by in vivo test using rodent models. One establishedmodel for peripherally elicited pruritus is through the injection ofserotonin into the rostral back area (neck) in hairless rats. Prior toserotonin injections (e.g., 2 mg/mL, 50 μL), a dose of a compound of thepresent invention can be applied systemically through oral, intravenousor intraperitoneal routes or topically to a circular area fixed diameter(e.g. 18 mm). Following dosing, the serotonin injections are given inthe area of the topical dosing. After serotonin injection the animalbehaviour is monitored by video recording for 20 min-1.5 h, and thenumber of scratches in this time compared to vehicle treated animals.Thus, application of a compound of the current invention could suppressserotonin-induced scratching in rats.

All of the U.S. patents, U.S. patent application publications, U.S.patent applications, foreign patents, foreign patent applications andnon patent publications referred to in this specification areincorporated herein by reference in their entireties.

Although the foregoing invention has been described in some detail tofacilitate understanding, it will be apparent that certain changes andmodifications may be practiced within the scope of the appended claims.Accordingly, the described embodiments are to be considered asillustrative and not restrictive, and the invention is not to be limitedto the details given herein, but may be modified within the scope andequivalents of the appended claims.

We claim:
 1. A compound of formula (I)

or a pharmaceutically acceptable salt thereof, wherein: A is a 6-12membered aryl ring that is optionally substituted with one or moregroups R^(A) independently selected from the group consisting of F, Cl,Br, I, —CN, —C(═O)OR^(A1), —SO₂R^(A1), —OR^(A1), —(X^(RA))—(3-15membered carbocyclyl), —(X^(RA))-(6 membered aryl), —(X^(RA))—(5-12membered heteroaryl), and —R^(A2), wherein said 3-15 memberedcarbocyclyl, 6-12 membered aryl, and 5-12 membered heteroaryl of R^(A)is optionally substituted with from 1 to 5 substituents R^(a)independently selected from the group consisting of F, Cl, Br, I,C₁₋₄alkyl, halo(C₁₋₄alkyl), amino, (C₁₋₄alkyl)amino, di(C₁₋₄alkyl)amino,halo(C₁₋₄alkoxy), and —C(O)N(R^(c))₂; R^(A1) is selected from the groupconsisting of hydrogen, C₁₋₈alkyl, C₂₋₈alkenyl, halo(C₁₋₈alkyl),C₃₋₈cycloalkyl, phenyl and benzyl; R^(A2) is selected from the groupconsisting of C₁₋₈alkyl that is optionally substituted with one or moresubstituents independently selected from oxo (=O), fluoro, hydroxy,amino, (C₁₋₄alkyl)amino and di(C₁₋₄alkyl)amino; X^(RA) is selected fromthe group consisting of absent, —C(═O)—, and C₁₋₄alkylene; wherein anyC₁₋₄alkylene, of X^(RA) is optionally substituted with 1 to 3substituents independently selected from the group consisting ofC₁₋₄alkyl, halo(C₁₋₄alkyl), and phenyl that is optionally substitutedwith 1 to 5 substituents independently selected from, F, Cl, Br, I,—NH₂, —OH, —CN, —NO₂, C₁₋₄alkyl, halo(C₁₋₄alkyl), C₁₋₄alkoxy,halo(C₁₋₄alkoxy), (C₁₋₄alkyl)amino and di(C₁₋₄alkyl)amino; B is a 5-12membered heteroaryl or a 6-12 membered aryl ring, wherein ring B isoptionally substituted with one or more groups R^(B) independentlyselected from the group consisting of F, Cl, Br, I, —CN, —C(═O)OR^(B1),—SO₂R^(B1), —OR^(B1), —(X^(RB))-(3-15 membered carbocyclyl),—(X^(RB))-(6-12 membered aryl), —(X^(RB))-(5-12 membered heteroaryl),and —R^(B2), wherein said 3-15 membered carbocyclyl, 6-12 membered aryl,and 5-12 membered heteroaryl of R^(B) is optionally substituted withfrom 1 to 5 substituents R^(b) independently selected from the groupconsisting of F, Cl, Br, I, C₁₋₄alkyl, halo(C₁₋₄alkyl), amino,(C₁₋₄alkyl)amino, di(C₁₋₄alkyl)amino, halo(C₁₋₄alkoxy), and—C(O)N(R^(C))₂; R^(B1) is selected from the group consisting ofhydrogen, C₁₋₈alkyl, C₂₋₈alkenyl, halo(C₁₋₈alkyl), C₃₋₈cycloalkyl,phenyl and benzyl; R^(B2) is selected from the group consisting ofC₁₋₈alkyl that is optionally substituted with one or more substituentsindependently selected from oxo (=O), fluoro, hydroxy, amino,(C₁₋₄alkyl)amino and di(C₁₋₄alkyl)amino; X^(RB) is selected from thegroup consisting of absent, —C(═O)—, and C₁₋₄alkylene; wherein anyC₁₋₄alkylene, of X^(RB) is optionally substituted with 1 to 3substituents independently selected from the group consisting ofC₁₋₄alkyl, halo(C₁₋₄alkyl), and phenyl that is optionally substitutedwith 1 to 5 substituents independently selected from, F, Cl, Br, I,—NH², —OH, —CN, —NO₂, C₁₋₄alkyl, halo(C₁₋₄alkyl), C₁₋₄alkoxy,halo(C₁₋₄alkoxy), (C₁₋₄alkyl)amino and di(C₁₋₄alkyl)amino; ring C is a5-, 6-, or 7-membered heterocyclyl, which 5-, 6-, or 7-memberedheterocyclyl is optionally substituted with one or more groups R^(c)independently selected from the group consisting of C1-4alkyl, halo, oxo(=O), thioxo (=S), and halo(C₁₋₄alkyl); R³, R⁴, and R⁵ are eachindependently selected from the group consisting of H, F, Cl, Br, I,—CN, C₁₋₈alkyl, C₃₋₈cycloalkyl, halo(C₁₋₄alkyl) and C₁₋₈alkoxy; and R⁶is selected from the group consisting of H, C₁₋₈alkyl, C₃₋₈ cycloalkyl,aryl, and aryl(C₁₋₄ alkyl); and R⁷ is selected from the group consistingof C₁₋₈alkyl, 3-15 membered heterocyclyl, 5-12 membered heteroaryl,—C(O)N(R^(d))₂ and —C(═NCN)N(R^(d))₂; or R⁶ and R⁷, together with thenitrogen to which they are attached, form a 3-15 membered heterocyclylor 5-12 membered heteroaryl; wherein any C₁₋₈ alkyl, C₃₋₈ cycloalkyl,aryl, aralkyl, 3-15 membered heterocyclyl, and 5-12 membered heteroarylis optionally substituted with one or more groups Re that areindependently selected from the group consisting of F, Cl, Br, I, —NH₂,—OH, —CN, —NO₂, C₁₋₄ alkyl, halo(C₁₋₄alkyl), C₁₋₄alkoxy,halo(C₁₋₄alkoxy), (C₁₋₄alkyl)amino and di(C₁₋₄alkyl)amino; each R^(d) isindependently selected from the group consisting of hydrogen, C₁₋₈alkyl, C₃₋₈ cycloalkyl, phenyl and benzyl.
 2. A compound of formula Ia:

or a pharmaceutically acceptable salt thereof, wherein: X is CH₂, O, S,SO, or SO₂; Y is H₂ or O; and Z is is CH₂ or O; provided that when Z isO, X is CH₂; A is a 6-12 membered aryl ring that is optionallysubstituted with one or more groups R^(A) independently selected fromthe group consisting of F, Cl, Br, I, —CN, —C(═O)OR^(A1), —SO₂R^(A1),—OR^(A1), —(X^(RA))—(3-15 membered carbocyclyl), —(X^(RA))—(6-12membered aryl), —(X^(RA))—(5-12) membered heteroaryl), and —R^(A2),wherein said 3-15 membered carbocyclyl, 6-12 membered aryl, and 5-12membered heteroaryl of R^(A) is optionally substituted with from 1 to 5substituents R^(a) independently selected from the group consisting ofF, Cl, Br, I, C₁₋₄ alkyl, halo(C₁₋₄alkyl), amino, (C₁₋₄alkyl)amino,di(C₁₋₄alkyl)amino, halo(C₁₋₄alkoxy), and —C(O)N(R^(c))₂; R^(A1) isselected from the group consisting of hydrogen, C₁₋₈ alkyl, C₂₋₈alkenyl, halo(C₁₋₈alkyl), C₃₋₈ cycloalkyl, phenyl and benzyl; R^(A2) isselected from the group consisting of C₁₋₈ alkyl that is optionallysubstituted with one or more substituents independently selected fromoxo (=O), fluoro, hydroxy, amino, (C₁₋₄alkyl)amino anddi(C₁₋₄alkyl)amino; X^(RA) is selected from the group consisting ofabsent, —C(═O)—, and C₁₋₄ alkylene; wherein any C₁₋₄alkylene, of X^(RA)is optionally substituted with 1 to 3 substituents independentlyselected from the group consisting of C₁₋₄ alkyl, halo(C₁₋₄ alkyl), andphenyl that is optionally substituted with 1 to 5 substituentsindependently selected from, F, Cl, Br, I, —NH₂, —OH, —CN, —NO₂, C₁₋₄alkyl, halo(C₁₋₄alkyl), C₁₋₄alkoxy, halo(C₁₋₄ alkoxy), (C₁₋₄ alkyl)aminoand di(C₁₋₄ alkyl)amino; B is a 5-12 membered heteroaryl or a 6-12membered aryl ring, wherein ring B is optionally substituted with one ormore groups R^(B) independently selected from the group consisting of F,Cl, Br, I, —CN, —C(═O)OR^(B1), —SO₂R^(B1), —OR^(B1), —(X^(RB))-(3-15membered carbocyclyl), —(X^(RB))-(6-12 membered aryl), —(X^(RB))-(5-12membered heteroaryl), and —R^(B2), wherein said 3-15 memberedcarbocyclyl, 6-12 membered aryl, and 5-12 membered heteroaryl of R^(B)is optionally substituted with from 1 to 5 substituents R^(b)independently selected from the group consisting of F, C₁, Br, I,C₁₋₄alkyl, halo(C₁₋₄alkyl), amino, (C₁₋₄alkyl)amino, di(Ci₁₋₄alkyl)amino, halo(C₁₋₈alkoxy), and —C(O)N(R^(c))₂, R^(B1) is selectedfrom the group consisting of hydrogen, C₁₋₈ alkyl, C₂₋₈ alkenyl,halo(C₁₋₈alkyl), C₃₋₈ cycloalkyl, phenyl and benzyl; R^(B2) is selectedfrom the group consisting of C₁₋₈ alkyl that is optionally substitutedwith one or more substituents independently selected from oxo (=O),fluoro, hydroxy, amino, (C₁₋₄ alkyl)amino and di(C₁₋₄ alkyl)amino;X^(RB) is selected from the group consisting of absent, —C(═O)—, andC₁₋₄ alkylene; wherein any C₁₋₄ alkylene, of X^(RB) is optionallysubstituted with 1 to 3 substituents independently selected from thegroup consisting of C₁₋₄ alkyl, halo(C₁₋₄alkyl), and phenyl that isoptionally substituted with 1 to 5 substituents independently selectedfrom, F, Cl, Br, I, —NH₂, —OH, —CN, —NO₂, C₁₋₄ alkyl, halo(C₁₋₄ alkyl),C₁₋₄ alkoxy, halo(C₁₋₄alkoxy), (C₁₋₄alkyl)amino and di(C₁₋₄alkyl)amino;R³, R⁴, and R⁵ are each independently selected from the group consistingof H, F, Cl, Br, I, —CN, C₁₋₈alkyl, C₃₋₈ cycloalkyl, halo(C₁₋₈alkyl) andC₁₋₈alkoxy; and R⁶ is selected from the group consisting of H, C₁₋₈alkyl, C₃₋₈ cycloalkyl, aryl, and aryl(C₁₋₄ alkyl); and R⁷ is selectedfrom the group consisting of C₁₋₈ alkyl, 3-15 membered heterocyclyl,5-12 membered heteroaryl, —C(O)N(R^(d))₂ and —C(═NCN)N(R^(d))₂; or R⁶and R⁷, together with the nitrogen to which they are attached, form a3-15 membered heterocyclyl or 5-12 membered heteroaryl; wherein anyC₁₋₈alkyl, C₃₋₈cycloalkyl, aryl, aralkyl, 3-15 membered heterocyclyl,and 5-12 membered heteroaryl is optionally substituted with one or moregroups R^(e) that are independently selected from the group consistingof F, Cl, Br, I, —NH₂, —OH, —CN, —NO₂, C₁₋₄ alkyl, halo(C₁₋₄alkyl),C₁₋₄alkoxy, halo(C₁₋₄ alkoxy), (C₁₋₄alkyl)amino and di(C₁₋₄ alkyl)amino;each R^(d) is independently selected from the group consisting ofhydrogen, C₁₋₈alkyl, C₃₋₈cycloalkyl, phenyl and benzyl.
 3. The compoundof claim 2, which is a compound of formula Ib:

wherein X is CH₂, O, S, SO, or SO₂; Y is H₂, or O; Z is CH₂ or O;provided that when Z is O, X is CH₂; and p is 1, 2, 3, or 4; or apharmaceutically acceptable salt thereof.
 4. The compound of claim 2,which is a compound of formula Ic:

wherein, X is CH₂, O, S, SO, or SO₂; Y is H₂, or O; Z is CH₂ or O;provided that when Z is O, X is CH₂; and p is 1, 2, 3, or 4; or apharmaceutically acceptable salt thereof.
 5. The compound of claim 2,which is a compound of formula Id:

wherein, X is CH₂, O, S, SO, or SO₂; Y is H₂, or O; Z is CH₂ or O;provided that when Z is O, X is CH₂; p is 1, 2, 3, or 4; and o is 1, 2,3, 4, or 5; or a pharmaceutically acceptable salt thereof.
 6. Thecompound of claim 2, which is a compound of formula Ie:

wherein Y is H₂ or O; p is 1, 2, 3, or 4; and o is 1, 2, 3, 4, or 5; ora pharmaceutically acceptable salt thereof.
 7. The compound of claim 2,which is a compound of formula If:

wherein, X is CH₂, O, S, SO, or SO₂; Y is H₂, or O; Z is CH₂ or O;provided that when Z is O, X is CH₂; and o is 1, 2, 3, 4, or 5; or apharmaceutically acceptable salt thereof.
 8. The compound of claim 2,which is a compound of formula Ig:

wherein Y is H₂ or O; and o is 1, 2, 3, 4, or 5; or a pharmaceuticallyacceptable salt thereof.
 9. The compound of claim 7, or apharmaceutically acceptable salt thereof, wherein X is O, S, SO, or SO₂;Y is H₂; and Z is CH₂.
 10. The compound of claim 7, or apharmaceutically acceptable salt thereof, wherein X is O, S, SO, or SO₂;Y is O; and Z is CH₂.
 11. The compound of claim 8, or a pharmaceuticallyacceptable salt thereof, wherein Y is O.
 12. The compound of claim 1, ora pharmaceutically acceptable salt thereof, wherein R³, R⁴, and R⁵ areeach independently selected from the group consisting of H, F, andC₁₋₄alkyl.
 13. The compound of claim 1, or a pharmaceutically acceptablesalt thereof, wherein —NR⁶R⁷ is:


14. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein —NR⁶R⁷ is:


15. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein R⁶ is H and R⁷ is a 5-6 membered heteroaryl.
 16. Thecompound of claim 1, or a pharmaceutically acceptable salt thereof,wherein each R^(A) is independently selected from the group consistingof F, Cl, trifluoromethoxy, CF₂, and CF₃.
 17. The compound of claim 1,or a pharmaceutically acceptable salt thereof, wherein each R^(B) isindependently selected from the group consisting of F, —CN, and methoxy.18. A compound selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.
 19. A pharmaceuticalcomposition comprising a compound of claim 1 or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable excipient.20. The compound of claim 1, wherein ring C is a 5-, 6-, or 7-memberedsaturated heterocyclyl, which 5-, 6-, or 7-membered saturatedheterocyclyl is optionally substituted with one or more groups R^(c)independently selected from the group consisting of C₁₋₄alkyl, halo, oxo(=O), thioxo (=S), and halo(C₁₋₄alkyl).